PONE-D-23-22896Evaluating possible maternal effect lethality and genetic background effects in Naa10 knockout micePLOS ONE

Dear Dr. LYON,

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Reviewers' comments:

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: Partly

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

Reviewer #1: No

Reviewer #2: Yes

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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

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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

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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 entitled „Evaluating possible maternal effect lethality and genetic background effects in Naa10 knockout mice“ is a well-designed and comprehensive analysis of a novel NAA10 inbred line generated in this study. The authors focus on replicating phenotypes described in previous studies and evaluating these effects. The results are well-described and clearly ordered. The authors could confirm the majority of previously described NAA10 impacts. The previously suggested "maternal effect lethality for heterozygous Naa10-/X female mice” was unreproducible, consistent with other authors' identifying a mice-specific NAA12 variant compensating for the loss of NAA10.

Minor concerns and suggestions for manuscript improvement

- The applied statistical tests are not mentioned in all cases. A section describing the statistical analyses is missing in the material and methods.

- PAGE 31 Pax3 mutant is not introduced. Please explain for the non-expert reader

- Page 31 “As such, there is modulation of these neural crest-related phenotypes by genetic background. There is a….” Rephrase these statements and the most relevant primary literature

- Discussion: The discussion could start with a broader perspective on NAA10 function in animals and other organisms (plants, fungi) and summarize the existence of NAA10 homologs in these species. This would help the non-expert readers to classify the here described findings in the context of NatA evolution.

Reviewer #2: In this manuscript, the authors tried to develop a mouse model for Ogden Syndrome with a Ser37Pro missense mutation by using two approaches. However, the gene editing did not work as they expected, and they obtained new knockout mice for Naa10. The authors found the new Naa10 knockout mice show similar phenotypes as previously reported (Kweon et al. 2021). Moreover, they demonstrated these phenotypes are modulated by genetic background and environmental effects. The reviewer has the following concerns for this manuscript.

Major concerns

1. In this manuscript, the authors observed embryonic lethality for some Naa10 knockout mice that were not reported in the previous publication (Kweon et al. 2021). The authors reasoned it is probably due to genetic backgrounds and/or environmental effects. The authors also stated it in the abstract as a new discovery for this manuscript. However, there are no data supporting this statement. The authors should perform some experiments such as examination of NatA enzyme activity to confirm NatA activity is influenced by the genetic backgrounds and environmental conditions in the new knockout mice.

2. The cardiac diseases are a common developmental anomaly in Ogden Syndrome patients. The abnormal cardiac development was reported in the previous Naa10 knockout mice (Kweon et al. 2021). The authors should examine the cardiac development in the new Naa10 knockout mice.

Minor concerns

In the Materials and Methods, the authors used GraphPad Prism 6 to plot the data for measurement of piebaldism. But in the Data Analysis, the authors stated Prism 9 Graphpad software version 9.5.1 was used. Why did the authors use two versions of the software?

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Reviewer #1: No

Reviewer #2: No

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Evaluating possible maternal effect lethality and genetic background effects in Naa10 knockout mice

PONE-D-23-22896R1

Dear Dr. LYON,

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.

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Michael Bader

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

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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: (No Response)

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

Reviewer #1: Yes

Reviewer #2: (No Response)

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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: (No Response)

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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: (No Response)

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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: The authors addressed all of my concerns. The revised manuscript is significantly improved and merits publication in PLOS One.

Reviewer #2: (No Response)

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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

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