Dear Dr. Mokdad-Gargouri,

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.

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Chunming Liu

Academic Editor

PLOS ONE

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Tunisian Ministry of Higher education and Scientific Research

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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?

Reviewer #1: Yes

Reviewer #2: Partly

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

Reviewer #1: Yes

Reviewer #2: N/A

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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: No

Reviewer #2: 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

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Reviewer #1: In this manuscript the authors confirmed 17 of 30 Tunisian colorectal cancer patients by identifying pathogenic variants using Sanger sequencing or Targeted NGS with a 31-gene panel. Results highlighted the necessity of genetic testing in confirming the genetic causes of CRC syndromes, especially in Lynch Syndrome. Minor issues:

1, Page 7 in the Results section, “five teen patients” should be fifteen patients.

2, Inconsistency of labeling: in figure 2a, “MP3” should be “MP13”. In addition, In page 7, patients were referred as MAP13, MAP14, MAP15 but in page 10 and in figure 2 they were labelled as MP13, MP14, MP15.

3, In page 5, the extra dot should be removed in “. Multiplex Ligation-Dependent Probe Amplification”.

4, The NGS sequencing data such as raw reads and SAM files should be deposited to a public repository.

5, Since this study focused on CRC samples, it would be better if authors discussed why the OncoRisk Cancer Panel was used instead of targeted CRC panel which typically includes other classical CRC oncogenes such as BRAF, KRAS and CTNNB1.

Reviewer #2: This study assessed the impact of genetic testing in the diagnosis of 30 patients suspected with hereditary polyposis or non-polyposis syndromes.

1. The authors should clarify the criteria/guidelines used to consider patients as suspected of having hereditary polyposis or non-polyposis syndromes. For example, did the authors follow NCCN Guidelines for Genetic/Familial High-Risk Assessment?

2. It would be helpful to include additional data on the concordance between Sanger sequencing and targeted NGS. Specifically, as both Sanger sequencing and targeted NGS were performed for patients MAP14 and MAP15, did the targeted NGS detect the same MUTYH variant identified by Sanger sequencing?

3. I couldn’t find Table 1 and Table 2.

4. The authors should carefully proofread the manuscript to correct typos. Below are a few noted:

Abstract, “hereditary polyposis syndromes such as MUTY-Associated Polyposis” should be “… MUTYH…”

Page 6, “. Multiplex Ligation-Dependent Probe Amplification” should be “Multiplex Ligation-Dependent Probe Amplification”

Page 7, “In this group, five teen patients suspected of MYH-Associated Polyposis” should be “… fifteen…”

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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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[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.

Genetic Profiling of Inherited Colorectal Cancer Syndromes in Tunisian Patients

PONE-D-25-11904R1

Dear Dr. Raja Mokdad-Gargouri,

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,

Chunming Liu

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

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??>

Reviewer #1: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: N/A

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

The PLOS Data policy

Reviewer #1: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: The authors addressed all my concerns.

Reviewer #2: The authors have addressed all my previous comments to a satisfactory level. I don’t have any further comments.

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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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