A single Musashi gene allele is sufficient to maintain mouse photoreceptor cells

Bohye Jeong; Peter Stoilov

doi:10.1371/journal.pone.0338576

Peer Review History

Original SubmissionNovember 24, 2025
13 Feb 2026 Decision Letter - Gerrit Hilgen, Editor -->PONE-D-25-63099-->-->A single Musashi gene allele is sufficient to maintain mouse photoreceptor cells-->-->PLOS One Dear Dr. Stoilov, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.-->--> Please submit your revised manuscript by Mar 30 2026 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. Please include the following items when submitting your revised manuscript:--> A letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols. We look forward to receiving your revised manuscript. Kind regards, Gerrit Hilgen Academic Editor PLOS One Journal Requirements: When submitting your revision, we need you to address these additional requirements. 1.Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. To comply with PLOS One submissions requirements, in your Methods section, please provide additional information regarding the experiments involving animals and ensure you have included details on (1) methods of sacrifice, (2) methods of anesthesia and/or analgesia, and (3) efforts to alleviate suffering. 3. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match. When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section. 4. Thank you for stating the following financial disclosure: “R01EY025536” Please state what role the funders took in the study. If the funders had no role, please state: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." If this statement is not correct you must amend it as needed. Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf. 5. We note that there is identifying data in the Supporting Information file “Supplementary_data_1_RawERG_data_and_R-code.zip” Due to the inclusion of these potentially identifying data, we have removed this file from your file inventory. Prior to sharing human research participant data, authors should consult with an ethics committee to ensure data are shared in accordance with participant consent and all applicable local laws. Data sharing should never compromise participant privacy. It is therefore not appropriate to publicly share personally identifiable data on human research participants. The following are examples of data that should not be shared: -Name, initials, physical address -Ages more specific than whole numbers -Internet protocol (IP) address -Specific dates (birth dates, death dates, examination dates, etc.) -Contact information such as phone number or email address -Location data -ID numbers that seem specific (long numbers, include initials, titled “Hospital ID”) rather than random (small numbers in numerical order) Data that are not directly identifying may also be inappropriate to share, as in combination they can become identifying. For example, data collected from a small group of participants, vulnerable populations, or private groups should not be shared if they involve indirect identifiers (such as sex, ethnicity, location, etc.) that may risk the identification of study participants. Additional guidance on preparing raw data for publication can be found in our Data Policy (https://journals.plos.org/plosone/s/data-availability#loc-human-research-participant-data-and-other-sensitive-data) and in the following article: http://www.bmj.com/content/340/bmj.c181.long. Please remove or anonymize all personal information (<specific identifying information in file to be removed>), ensure that the data shared are in accordance with participant consent, and re-upload a fully anonymized data set. Please note that spreadsheet columns with personal information must be removed and not hidden as all hidden columns will appear in the published file. 6. Please include your full ethics statement in the ‘Methods’ section of your manuscript file. In your statement, please include the full name of the IRB or ethics committee who approved or waived your study, as well as whether or not you obtained informed written or verbal consent. If consent was waived for your study, please include this information in your statement as well. 7. PLOS ONE now requires that authors provide the original uncropped and unadjusted images underlying all blot or gel results reported in a submission’s figures or Supporting Information files. This policy and the journal’s other requirements for blot/gel reporting and figure preparation are described in detail at https://journals.plos.org/plosone/s/figures#loc-blot-and-gel-reporting-requirements and https://journals.plos.org/plosone/s/figures#loc-preparing-figures-from-image-files. When you submit your revised manuscript, please ensure that your figures adhere fully to these guidelines and provide the original underlying images for all blot or gel data reported in your submission. See the following link for instructions on providing the original image data: https://journals.plos.org/plosone/s/figures#loc-original-images-for-blots-and-gels. In your cover letter, please note whether your blot/gel image data are in Supporting Information or posted at a public data repository, provide the repository URL if relevant, and provide specific details as to which raw blot/gel images, if any, are not available. Email us at plosone@plos.org if you have any questions. 8. Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information. 9. If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. Additional Editor Comments: Dear authors, The reviewers would like to see a bit more extended discussion and more details about cell types and methodology. Most comments can be implemented on the existing data. However, I don't see a need to do more experiments. Please see the review comments for details. I am looking forward seeing your revised manuscript [Note: HTML markup is below. Please do not edit.] Reviewer's Responses to Questions -->Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. --> Reviewer #1: Yes Reviewer #2: Yes ******** -->2. Has the statistical analysis been performed appropriately and rigorously? --> Reviewer #1: Yes Reviewer #2: Yes ****** -->3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.--> Reviewer #1: Yes Reviewer #2: Yes ****** -->4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.--> Reviewer #1: Yes Reviewer #2: Yes ****** -->5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)--> Reviewer #1: The manuscript by Jeong and Stoilov, "A Single Musashi Gene Allele Is Sufficient to Maintain Mouse Photoreceptor Cells," appears to be a continuation of the research being conducted by this group and is devoted to studying how many alleles of the Msi1 and Msi2 genes are required to maintain the normal structure and function of photoreceptors. The manuscript is written in good scientific English, the rationale for the research question is clearly presented, and the materials and methods are described accurately and thoroughly. The study was conducted at a high methodological level. The manuscript is sufficiently well illustrated to ensure the reliability of the results obtained. However, I have several recommendations/comments regarding the manuscript text that would be worth considering before it is published: 1) Since the authors recently published a paper on a very similar topic (The Musashi proteins MSI1 and MSI2 are required for photoreceptor morphogenesis and vision in mice, 2020), I would like to see a more detailed discussion in the introduction of what is new in the current manuscript, which is the reason for this clarifying work. 2) I would like to understand the basis for choosing the time points for performing different analyses after tamoxifen injections (e.g., 14 days for genotyping, 143 days for collecting retinas for immunohistochemistry). This should either be supported by some references, or you can cite your own experience. 3) Clarification is needed in the Materials and Methods section regarding what TUBB is and why it is used as a loading control. Some references are needed on where this approach was borrowed. 4) In the subsection 'A single allele of Musashi is sufficient to maintain photoreceptor cell function' in the Results section, was the ERG response examined to only one intensity? Figure 3 appears to show only one stimulus intensity. If so, how was this particular stimulus intensity chosen? 5) In this subsection, it is useful to emphasize that both photopic and scotopic ERGs were recorded, allowing us to differentiate between the preservation of rod and cone function. 6) Figure 3 – the current design is poor and unreadable. It has too many elements, and the animal group labels and axes are excessively small. It is necessary to consider the reader's needs and find a solution to make this figure more readable. 7) Caption to Figure 3, panels B and C (lines 262-263) – I strongly recommend replacing the word 'intensity' with 'amplitude'; this would be more terminologically correct. Reviewer #2: In this manuscript, Jeong et Stoilov investigate the role of the Musashi genes, Msi1 and Msi2, in regulating photoreceptor-specific alternative splicing. By progressively reducing Musashi gene dosage in photoreceptors, the authors assess how allele number affects photoreceptor genes and photoreceptor function. Their results indicate that even a single Msi1 or Msi2 allele is sufficient to preserve splicing of photoreceptor-specific exons and maintain photoreceptor function. The manuscript is clearly written, well-illustrated, and overall easy to follow. The study is interesting and the data are potentially valuable. However, several points should be clarified or addressed before this manuscript can be considered for publication. Major comments: • The authors should clearly indicate which retinal cell types express the Cre recombinase in the Pde6g-CreERT2 line. Is Cre activity restricted to rods, or present in both rods and cones? • The authors state that Western blot confirmed “expected changes” in Msi1 and Msi2 levels. Please specify what those expected changes are. When only one allele is missing, it is difficult to appreciate whether the other allele leads to increased protein expression. This point should be developed. • Among the genes tested for differential splicing, how many splice variants were covered by the primers? Could alternative splicing events occur due to imbalance of Msi1 / Msi2 levels? Did the authors also measure total gene expression levels, not only splice variants? • Did the authors examine other splicing factors that might compensate via increased expression (e.g., PRPF family members)? • Additional immunohistochemistry would strengthen the study, particularly for cilia and photoreceptor markers. GFAP staining would help assess retinal stress and Müller glia activation. Lack of GFAP upregulation would support absence of phenotype. Minor comments: • The text suggests a single intraperitoneal tamoxifen injection (p.5), but the Western blot section states that retinas were collected 14 days after the first tamoxifen injection. This implies that multiple injections may have been performed. Please clarify. • Were splicing changes assessed at later time points? A compensatory effect could diminish over time, so longer follow-up would be informative. • The schematic is helpful for readers. It might be easier to follow if placed between the MSI1 and MSI2 gels. • Please verify whether the correct citation is Sundar et al., 2021 rather than Matalkah et al., 2022. • One hundred days may be insufficient to detect subtle or slowly progressive defects. This limitation should be acknowledged. • The legend states a 40× objective, but the images look closer to 20×. Please verify. In addition, a higher magnification of ONL/IS area in Fig. 4C should be added to better illustrate the findings. • Can the authors comment on the strong increase of MSI1 or MSI2 in the INL when the other gene is completely missing? • The authors are thanked for sharing their R scripts, but they cannot be fully tested because the file Light_intensity_calibration.csv is missing. ****** -->6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.--> Reviewer #1: No Reviewer #2: No ******** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] To ensure your figures meet our technical requirements, please review our figure guidelines: https://journals.plos.org/plosone/s/figures You may also use PLOS’s free figure tool, NAAS, to help you prepare publication quality figures: https://journals.plos.org/plosone/s/figures#loc-tools-for-figure-preparation. NAAS will assess whether your figures meet our technical requirements by comparing each figure against our figure specifications. https://doi.org/10.1371/journal.pone.0338576.r001
Revision 1
6 Apr 2026 Author Response We would like to thank the reviewers for devoting time to read our manuscript and providing thorough and detailed critiques. Our responses to the specific issues are below. Reviewer #1: 1) Since the authors recently published a paper on a very similar topic (The Musashi proteins MSI1 and MSI2 are required for photoreceptor morphogenesis and vision in mice, 2020), I would like to see a more detailed discussion in the introduction of what is new in the current manuscript, which is the reason for this clarifying work. The rationale behind this study is not to demonstrate that the Musashi proteins are required for vision. As the reviewer points out we have already shown this (Sundar et al 2020, PMID: 33168629 and Matalkah et al 2022, PMID: 36153373). The rationale is related to the role of Musashi in regulating splicing specifically in photoreceptor cells, which is a nuclear function for the two proteins. It was previously shown that the nuclear localization signal of the Musashi proteins is on their RNA binding surface, which makes RNA binding and nuclear import mutually exclusive. In this paper we were testing the hypothesis that the Musashi proteins regulate splicing specifically in photoreceptor cells due to their high level of expression in this cell type. The logic was that the high levels of Musashi proteins saturate their cytoplasmic mRNA targets and the excess free protein can be imported into the nucleus to regulate splicing. We have edited the introduction (lines 58-60 in the track changes version of the revised manuscript) to make that point more clear. 2) I would like to understand the basis for choosing the time points for performing different analyses after tamoxifen injections (e.g., 14 days for genotyping, 143 days for collecting retinas for immunohistochemistry). This should either be supported by some references, or you can cite your own experience. There must be some misunderstanding due to us combining the genotyping results and western blot analysis of the knockouts into one. We genotype the animals at weaning (post natal day 21). The knockout is induced at postnatal day 30 with injection of tamoxifen. The western blot analysis was performed 14 days later. The 14 day timepoint is based on our previous work (Matalkah et al 2022, PMID: 36153373), where we did a time course of Musashi protein levels and showed that two weeks after the knockout the protein levels reached their minimum. We amended the text (lines 233-236 in the track changes version of the revised manuscript) to clarify that point. To reduce animal usage, eye cups for MSI1 and MSI2 immunofluorescence imaging (Figure 4) were from the animals that were used for ERG experiments. The ERG experiments were completed 112 days post tamoxifen injections and analysis took some additional time. This pushed the eye cup collection to 143 days post tamoxifen injection. We clarified this point on line 132 in the Materials and Methods section in the track changes version of the revised manuscript) 3) Clarification is needed in the Materials and Methods section regarding what TUBB is and why it is used as a loading control. Some references are needed on where this approach was borrowed. TUBB refers to beta tubulin as recognized by the Sigma-Aldrich antibody T8328 (listed in Supplementary table 2). Beta tubulins are abundant cytoskeletal proteins in neural tissues. Beta tubulin antibodies are commonly used as loading controls, and we have used them as such in our previous studies of the Musashi proteins in mouse retina. While we could have used other loading controls or normalized to total protein, use of beta tubulin provides consistency with our previous research. We have amended the Materials and Methods section on lines 125-127 in the track changes version of the revised manuscript to clarify this point. 4) In the subsection 'A single allele of Musashi is sufficient to maintain photoreceptor cell function' in the Results section, was the ERG response examined to only one intensity? Figure 3 appears to show only one stimulus intensity. If so, how was this particular stimulus intensity chosen? We routinely do our ERG experiments using four intensities ( -40 dB, -24 dB, -12 dB, -4 dB) to assess scotopic response, and 3 dB for photopic response. -12 dB intensity was chosen because it elicits strong rod photoreceptor response, without activating cones. We have added a supplementary figure 4B to show the scotopic intensity-response curves. 5) In this subsection, it is useful to emphasize that both photopic and scotopic ERGs were recorded, allowing us to differentiate between the preservation of rod and cone function. We addressed this concern in lines 272-273 of the Results section in the track changes version of the revised manuscript. 6) Figure 3 – the current design is poor and unreadable. It has too many elements, and the animal group labels and axes are excessively small. It is necessary to consider the reader's needs and find a solution to make this figure more readable. To make the figure 3 readable we moved panel A to supplementary figure 4A and enlarged the legend and labels text. 7) Caption to Figure 3, panels B and C (lines 262-263) – I strongly recommend replacing the word 'intensity' with 'amplitude'; this would be more terminologically correct. We apologize for this error. Changed as suggested. Reviewer #2: Major comments: • The authors should clearly indicate which retinal cell types express the Cre recombinase in the Pde6g-CreERT2 line. Is Cre activity restricted to rods, or present in both rods and cones? While the Pde6g gene encodes the rod cGMP phosphodiesterase, it is expressed in cones and can compensate for the loss of the Pde6h gene that encodes the cone phosphodiesterase enzyme (Brennenstuhl et al 2015, PMID: 25739440). Consequently the Pde6g-CreERT2 knock-in line expresses Cre in both rod and cone photoreceptors. This is also evident from the immunofluorescence staining on Figure 4, where there are no cells in the photoreceptor layer that express MSI1 or MSI2 protein when both alleles of the respective gene are targeted. At the same time cones are readily detectable by PNA staining (Figure 5). We have previously shown (Matalkah et al 2022, PMID: 36153373) that deleting both MSI1 and MSI2 using Ped6g-CreERT2 abolishes both cone and rod photoreceptor response to light. • The authors state that Western blot confirmed “expected changes” in Msi1 and Msi2 levels. Please specify what those expected changes are. When only one allele is missing, it is difficult to appreciate whether the other allele leads to increased protein expression. This point should be developed. The expected change was 50-70% reduction of MSI1 or MSI2 protein in retinal lysates when both alleles of the respective genes are targeted in photoreceptor cells. As suggested we elaborated this point on lines 228-231 of the results section in the track changes version of the revised manuscript. • Among the genes tested for differential splicing, how many splice variants were covered by the primers? Could alternative splicing events occur due to imbalance of Msi1 / Msi2 levels? Did the authors also measure total gene expression levels, not only splice variants? Primers used for alternative splicing analysis on figure 2 are placed in the constitutive exons flanking a single alternative cassette exon. Each primer set covers the splicing of one alternative exon and detects two splice variants - one that includes and one that skips the exon. We have added “+” and “-” labels to the gel images in Figure 2 to mark the exon included and exon skipped variants. These splice variants should not be confused with the transcript variants produced by the gene - a gene can produce more than one transcript variant can include a particular alternative exon, depending on the splicing of other alternative exons in the same gene, and the use of alternative promoters and poly-adenylation sites. We are not sure what exactly the reviewer means by “imbalance of Msi1 / Msi2 levels”. We can think of two interpretations - either different ratios of MSI1 to MSI2 protein, or differences in the combined expression of MSI1 and MSI2. We do not find a significant difference in the inclusion of the tested exons when only a single MSI1 allele is present compared to when only a single MSI2 allele is present, nor do we see any difference in splicing across genotypes that delete two alleles, regardless to which gene these alleles belong. This data would argue that the splicing is not affected by the ratio of MSI1 to MSI2. If we consider the combined MSI1 and MSI2 expression, loss of both proteins significantly impacts the splicing of the tested exons. In all cases the presence of a single Musashi allele is sufficient to promote the splicing of the tested exons to levels that were comparable to the controls. We did not analyze total gene expression levels in the Musashi allele knockouts used in this work. We previously analyzed by RNA-Seq the transcript levels in the double Msi1/Msi2 knockout generated using identical Pde6G-CreERT2 induced knockout. In these experiments we did not observe significant changes in the transcript levels of Prom1, Cc2d2a, Cep290, and Ttc8 (Matalkah et al 2022, PMID: 36153373 - see Supplementary data 1 and 8 in that publication). Therefore we do not expect to see significant transcript changes in knockouts that preserve one or more of the Musashi alleles • Did the authors examine other splicing factors that might compensate via increased expression (e.g., PRPF family members)? We did not examine the levels of other splicing factors in this work, however we did that previously for the complete Msi1/Msi2 knockout in (Matalkah et al 2022, PMID: 36153373 - supplementary data 1). There were no significant changes in splicing factor transcript levels. Neither transcript nor protein levels of Prpf or other core spliceosome proteins were affected in the Msi1/Msi2 knockout. However, SRSF9 protein levels as detected by mass spec were significantly changed (1.7 fold increase adjusted p-value < 0.001; Matalkah et al 2022, PMID: 36153373 - supplementary data 5). The change in the protein levels of SRSF9 appears to be a result of increased skipping of its poison exon in the Musashi knockout. • Additional immunohistochemistry would strengthen the study, particularly for cilia and photoreceptor markers. GFAP staining would help assess retinal stress and Müller glia activation. Lack of GFAP upregulation would support absence of phenotype. We thank the reviewer for the suggestion. We performed GFAP staining on retinal sections from mice expressing single Msi1 or Msi2 allele (Figure 6). We do not see elevated GFAP expression in Müller glia from the retina of animals expressing a single Musashi allele. Astrocyte projections in the ganglion cell layer stained for GFAP and served as internal positive control. Minor comments: • The text suggests a single intraperitoneal tamoxifen injection (p.5), but the Western blot section states that retinas were collected 14 days after the first tamoxifen injection. This implies that multiple injections may have been performed. Please clarify. Tamoxifen was injected on three consecutive days starting at postnatal day 30. Day count starts from the day of the first injection in the series. We apologize for the confusion - the materials and methods section for the tamoxifen injections was not clearly written. We have corrected the text of this section (lines 105-113 in the track changes version of the revised manuscript). • Were splicing changes assessed at later time points? A compensatory effect could diminish over time, so longer follow-up would be informative. We agree that compensatory effects can change over time. We have not assessed spicing changes at a later point. Due to the large number of genotypes we had to make a choice on how we allocate the samples for each experiment. Our choice was to use the longer time point samples for immunofluorescence. We have acknowledged the limited time course during which the phenotype of the animals was tracked as a limitation of the study (lines 406-409 in the track changes version of the revised manuscript). • The schematic is helpful for readers. It might be easier to follow if placed between the MSI1 and MSI2 gels. We tried to reformat Figure 1 as suggested, but this made the figure less readable for us as it spaced the gels apart. We prefer the keep in its current format. • Please verify whether the correct citation is Sundar et al., 2021 rather than Matalkah et al., 2022. We are not sure to which specific citation the reviewer refers to. After reviewing the citations we noticed one discrepancy where both Sundar and Matalkah were referenced regarding the inducible photoreceptor knockouts (line 223 in the track changes version of the revised manuscript) when only Matalkah was relevant. We have corrected this citation. • One hundred days may be insufficient to detect subtle or slowly progressive defects. This limitation should be acknowledged. We agree and we have acknowledged this limitation in the discussion (lines 406-409 in the track changes version of the revised manuscript). • The legend states a 40× objective, but the images look closer to 20×. Please verify. In addition, a higher magnification of ONL/IS area in Fig. 4C should be added to better illustrate the findings. We have used 40x objectives to image these sections. The image may appear as lower magnification because we used a larger field of view to show a broad section of the retina. We have added cutouts at larger magnification in supplementary figures 5 and 6. • Can the authors comment on the strong increase of MSI1 or MSI2 in the INL when the other gene is completely missing? This is likely due to variation of the staining across samples and replicates. This can be observed on supplementary figure 7 where the staining intensity of INL varies across replicates of the same genotype tracking the overall brightness of the image. • The authors are thanked for sharing their R scripts, but they cannot be fully tested because the file Light_intensity_calibration.csv is missing. We apologize for the omission. We have added the missing file to the archive. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0338576.r002
26 Apr 2026 Decision Letter - Gerrit Hilgen, Editor <p>A single Musashi gene allele is sufficient to maintain mouse photoreceptor cells PONE-D-25-63099R1 Dear Dr. Stoilov, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact billing support. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Gerrit Hilgen Academic Editor PLOS One Additional Editor Comments (optional): Reviewers' comments: Reviewer's Responses to Questions -->Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.--> Reviewer #1: All comments have been addressed Reviewer #2: All comments have been addressed ******** -->2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. --> Reviewer #1: Yes Reviewer #2: Yes ****** -->3. Has the statistical analysis been performed appropriately and rigorously? --> Reviewer #1: Yes Reviewer #2: Yes ****** -->4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.--> Reviewer #1: Yes Reviewer #2: Yes ****** -->5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.--> Reviewer #1: Yes Reviewer #2: Yes ****** -->6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)--> Reviewer #1: Even before the review, the work was well prepared and presented, and now the authors have adequately addressed the minor issues I pointed out. I am satisfied and believe the manuscript can be accepted for publication in its current form. Reviewer #2: Most of my comments have been carefully addressed by the authors, and the manuscript has improved accordingly. The revisions have clarified several important points and strengthened the overall quality of the work. In its current form, I consider the manuscript suitable for publication. ****** -->7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.--> Reviewer #1: No Reviewer #2: No ******** https://doi.org/10.1371/journal.pone.0338576.r003
Formally Accepted
Acceptance Letter - Gerrit Hilgen, Editor PONE-D-25-63099R1 PLOS One Dear Dr. Stoilov, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS One. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps. Lastly, if your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. If we can help with anything else, please email us at customercare@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Gerrit Hilgen Academic Editor PLOS One https://doi.org/10.1371/journal.pone.0338576.r004

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .