Dear Dr. Vo,

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 May 12 2025 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.

• A rebuttal 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'.
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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.

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We look forward to receiving your revised manuscript.

Kind regards,

Chen Ling, Ph.D.

Academic Editor

PLOS ONE

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Additional Editor Comments (if provided):

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Yes

Reviewer #2: Partly

Reviewer #3: Yes

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2. Has the statistical analysis been performed appropriately and rigorously? -->?>

Reviewer #1: N/A

Reviewer #2: N/A

Reviewer #3: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available??>

The PLOS Data policy

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English??>

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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Reviewer #1: 1. The author should add the full name before the first use of a certain abbreviation�including “CRISPR-Cas9”�. And the Introduction part should include general introduction�history, background�, why to write this review, the focus and highlights of this review, the following logical sequence of the review, best presented in 3-5 paragraphs.

2. Considering that most of the included studies used the CRISPR system, I suggest the author add a separate section to summarize the discovery and development of the basic CRISPR system and base editors.

3. I understand that the authors‘ main focus is on screened articles, but the authors mention the use of gene editing therapies in animal models in the title. I would suggest that the authors add a separate section to discuss the impact of various types of animal models in gene editing therapies for cardiovascular disease, especially given that some studies are currently using rabbits, dogs and cats, among others.

4. Ethical issues of gene editing treatments should be mentioned.

5. AAV subtypes (AAV8, AAV9) should be discussed.

Reviewer #2: I like the topic of this review. If it could be properly written, it will be an important reference for the field. However, there are major problems in the contents and logics. The following questions need to be solved:

1. There must be something wrong with keyword search. You need to truly understand this field in order to write a good review paper. You need to curate literature search manually since some good papers could not be easily searched by just looking at title or abstract. Just to give you some examples of missed key papers in high-profile journals:

� Lebek S, Chemello F, Caravia XM, Tan W, Li H, Chen K, Xu L, Liu N, Bassel-Duby R, Olson EN. Ablation of CaMKIIδ oxidation by CRISPR-Cas9 base editing as a therapy for cardiac disease. Science. 2023 Jan 13;379(6628):179-185. doi: 10.1126/science.ade1105. Epub 2023 Jan 12. PMID: 36634166; PMCID: PMC10150399.

� Lebek S, Caravia XM, Chemello F, Tan W, McAnally JR, Chen K, Xu L, Liu N, Bassel-Duby R, Olson EN. Elimination of CaMKIIδ Autophosphorylation by CRISPR-Cas9 Base Editing Improves Survival and Cardiac Function in Heart Failure in Mice. Circulation. 2023 Nov 7;148(19):1490-1504. doi: 10.1161/CIRCULATIONAHA.123.065117. Epub 2023 Sep 15. PMID: 37712250; PMCID: PMC10842988.

� Wu S, Yang P, Geng Z, Li Y, Guo Z, Lou Y, Zhang S, Xiong J, Hu H, Guo X, Pu WT, Zhang Y, Zhu D, Zhang B. Base editing effectively prevents early-onset severe cardiomyopathy in Mybpc3 mutant mice. Cell Res. 2024 Apr;34(4):327-330. doi: 10.1038/s41422-024-00930-7. Epub 2024 Feb 9. PMID: 38337022; PMCID: PMC10978934.

� Liu Z, Yang L, Yang Y, Li J, Chen Z, Guo C, Guo Q, Li Q, Zhao D, Hu X, Gao F, Guo Y. ABE-Mediated Cardiac Gene Silencing via Single AAVs Requires DNA Accessibility. Circ Res. 2025 Jan 31;136(3):318-320. doi: 10.1161/CIRCRESAHA.124.325611. Epub 2025 Jan 16. PMID: 39817340.

� Yang L, Liu Z, Chen G, Chen Z, Guo C, Ji X, Cui Q, Sun Y, Hu X, Zheng Y, Li Y, Gao F, Chen L, Zhou P, Pu WT, Guo Y. MicroRNA-122-Mediated Liver Detargeting Enhances the Tissue Specificity of Cardiac Genome Editing. Circulation. 2024 May 28;149(22):1778-1781. doi: 10.1161/CIRCULATIONAHA.123.065438. Epub 2024 May 28. PMID: 38805581.

� Ghahremani S, Kanwal A, Pettinato A, Ladha F, Legere N, Thakar K, Zhu Y, Tjong H, Wilderman A, Stump WT, Greenberg L, Greenberg MJ, Cotney J, Wei CL, Hinson JT. CRISPR Activation Reverses Haploinsufficiency and Functional Deficits Caused by TTN Truncation Variants. Circulation. 2024 Apr 16;149(16):1285-1297. doi: 10.1161/CIRCULATIONAHA.123.063972. Epub 2024 Jan 18. PMID: 38235591; PMCID: PMC11031707.

� Ma S, Jiang W, Liu X, Lu WJ, Qi T, Wei J, Wu F, Chang Y, Zhang S, Song Y, Bai R, Wang J, Lee AS, Zhang H, Wang Y, Lan F. Efficient Correction of a Hypertrophic Cardiomyopathy Mutation by ABEmax-NG. Circ Res. 2021 Oct 29;129(10):895-908. doi: 10.1161/CIRCRESAHA.120.318674. Epub 2021 Sep 16. PMID: 34525843.

2. the paper need to be rewriten to discuss some key concepts with better structures and logics:

� AAV vs LNP vs AdV: what are differences between these vectors? The Pros and Cons of each of these vectors should be analysed and described. Why some studies used AAV while other studies used LNP? What is the rationale behind vector choices?

� Heart vs Liver: why some studies target the liver while other studies target the heart? Are they treating different type of diseases? What is the rationale behind organ choices? Are there difficulties or concerns to deliver to each organ?

� Cas9 NHEJ vs base editing vs epigenome editing vs RNA editing vs HDR: the paper should explain working mechanisms of each editor and their strength or weakness. Why certain studies used certain editor?

� Rare disease versus common disease: some therapies are specific for rare disease only (such as ABE correction of rare disease mutation). Other therapies are suitable for both rare and common diseases (such as pcsk9 and camk2d studies). Analysis from this angle is necessary to determine which type of therapy is more realistic and promising.

� Safety concerns from vector, editor, gene target should be separately and more detailed and systemically summarized.

Reviewer #3: This review examines the application of gene editing technologies in animal models of CVDs, summarizing the therapeutic efficacy and safety profiles of tools such as CRISPR-Cas9 in treating hyperlipidemia, cardiomyopathy, and related conditions. The article also discusses current limitations and future directions, including clinical translation and ethical considerations. While the manuscript presents a logically structured and comprehensive overview, certain aspects require supplementation.

1. The Introduction fails to explicitly delineate the fundamental distinctions between gene editing and conventional therapies (e.g., target specificity). The latest studies such as PMID: 37662968 can be referenced.

2. Page 8 states "Lipid nanoparticles were employed...to the liver (PCSK9, LDLR)" without explaining why the liver serves as a critical therapeutic target for CVD treatment, creating an incomplete logical progression.

3. Page 8 notes "editing efficiency varies across species" but lacks mechanistic analysis of underlying causes.

4. Page 14 could benefit from incorporation of recent findings (e.g., PMID: 40061823).

5. Page 18 requires more detailed technical descriptions of gene editing protocols, particularly CRISPR-Cas9 sgRNA design principles.

6. Page 11 should address interspecies biological differences in gene editing responses (e.g., mice vs. non-human primates) and their clinical translation implications.

7. Pages 20-23 would be strengthened by including updates on human clinical trials, particularly late-stage or approved therapies.

8. The verbose sentence on Page 4 ("Early gene replacement therapies...") could be condensed to: "Early therapies introduced functional gene copies to restore normal function."

9. In Introduction or Discussion, current advances in AAV research (e.g., transduction efficiency enhancement) should be cited (e.g., PMID: 38307819).

10. Terminology inconsistency ("CRISPR-Cas9" vs. "CRISPR/Cas9") requires standardization.

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Reviewer #1: No

Reviewer #2: Yes:  Yuxuan Guo

Reviewer #3: 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.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step.

Dear Dr. Vo,

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 Jun 14 2025 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.

• A rebuttal 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,

Chen Ling, Ph.D.

Academic Editor

PLOS ONE

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #2: (No Response)

Reviewer #3: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions??>

Reviewer #2: Yes

Reviewer #3: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously? -->?>

Reviewer #2: N/A

Reviewer #3: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available??>

The PLOS Data policy

Reviewer #2: Yes

Reviewer #3: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English??>

Reviewer #2: Yes

Reviewer #3: Yes

**********

Reviewer #2: The authors addressed most of my questions properly, except for the exclusion of important short-format papers that are still quite important to the field. Research letters are still original research articles, not simple correspondences.

1. Thank you for highlighting these omissions and providing valuable references. I would like to clarify that some of the suggested articles (e.g., doi: 10.1038/s41422-024-00930-7, 10.1161/CIRCRESAHA.124.325611, and 10.1161/CIRCULATIONAHA.123.065438) were identified during my initial search but were excluded as they are letters or correspondences, which did not meet our inclusion criteria focused on original research.

Re: This is exactly the problem with your exclusion criteria. These excluded research letter papers are still original research and they are peer-reviewed following the same standard as research articles. Their shorter formats simply mean they are focused on single strong points that do not need longer formats. You should still include these papers in your review.

Reviewer #3: The author has carefully revised the manuscript according to the comments of the reviewer, and there is no problem now. It is recommended to accept.

**********

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 #2: Yes:  Yuxuan Guo

Reviewer #3: 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.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step.

Gene editing therapy as a therapeutic approach for cardiovascular diseases in animal models: a scoping review

PONE-D-25-07256R2

Dear Dr. Vo,

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.

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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,

Chen Ling, Ph.D.

Academic Editor

PLOS ONE

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #2: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions??>

Reviewer #2: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously? -->?>

Reviewer #2: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available??>

The PLOS Data policy

Reviewer #2: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English??>

Reviewer #2: Yes

**********

Reviewer #2: All my concerns have been properly addressed. I have no further comments on the content of this manuscript.

**********

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 #2: No

**********