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Combined Use of Whole Exome Sequencing and CRISPR/Cas9 to Study the Etiology of Non-Obstructive Azoospermia: Demonstration of the Dispensable Role of the Testis-Specific Genes C1orf185 and CCT6B

The genetic landscape of male infertility is highly complex. It is estimated that at least 4000 genes are involved in human spermatogenesis, but only few have so far been extensively studied. In this study, we investigated by whole exome sequencing two cases of idiopathic non-obstructive azoospermia...

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Autores principales: Cazin, Caroline, Neirijnck, Yasmine, Loeuillet, Corinne, Wehrli, Lydia, Kühne, Françoise, Lordey, Isabelle, Mustapha, Selima Fourati Ben, Bouker, Amin, Zouari, Raoudha, Thierry-Mieg, Nicolas, Nef, Serge, Arnoult, Christophe, Ray, Pierre F., Kherraf, Zine-Eddine
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8750304/
https://www.ncbi.nlm.nih.gov/pubmed/35011680
http://dx.doi.org/10.3390/cells11010118
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author Cazin, Caroline
Neirijnck, Yasmine
Loeuillet, Corinne
Wehrli, Lydia
Kühne, Françoise
Lordey, Isabelle
Mustapha, Selima Fourati Ben
Bouker, Amin
Zouari, Raoudha
Thierry-Mieg, Nicolas
Nef, Serge
Arnoult, Christophe
Ray, Pierre F.
Kherraf, Zine-Eddine
author_facet Cazin, Caroline
Neirijnck, Yasmine
Loeuillet, Corinne
Wehrli, Lydia
Kühne, Françoise
Lordey, Isabelle
Mustapha, Selima Fourati Ben
Bouker, Amin
Zouari, Raoudha
Thierry-Mieg, Nicolas
Nef, Serge
Arnoult, Christophe
Ray, Pierre F.
Kherraf, Zine-Eddine
author_sort Cazin, Caroline
collection PubMed
description The genetic landscape of male infertility is highly complex. It is estimated that at least 4000 genes are involved in human spermatogenesis, but only few have so far been extensively studied. In this study, we investigated by whole exome sequencing two cases of idiopathic non-obstructive azoospermia (NOA) due to severe hypospermatogenesis. After variant filtering and prioritizing, we retained for each patient a homozygous loss-of-function (LoF) variant in a testis-specific gene, C1orf185 (c.250C>T; p.Gln84Ter) and CCT6B (c.615-2A>G), respectively. Both variants are rare according to the gnomAD database and absent from our local control cohort (n = 445). To verify the implication of these candidate genes in NOA, we used the CRISPR/Cas9 system to invalidate the mouse orthologs 4930522H14Rik and Cct6b and produced two knockout (KO) mouse lines. Sperm and testis parameters of homozygous KO adult male mice were analyzed and compared with those of wild-type animals. We showed that homozygous KO males were fertile and displayed normal sperm parameters and a functional spermatogenesis. Overall, these results demonstrate that not all genes highly and specifically expressed in the testes are essential for spermatogenesis, and in particular, we conclude that bi-allelic variants of C1orf185 and CCT6B are most likely not to be involved in NOA and male fertility.
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spelling pubmed-87503042022-01-12 Combined Use of Whole Exome Sequencing and CRISPR/Cas9 to Study the Etiology of Non-Obstructive Azoospermia: Demonstration of the Dispensable Role of the Testis-Specific Genes C1orf185 and CCT6B Cazin, Caroline Neirijnck, Yasmine Loeuillet, Corinne Wehrli, Lydia Kühne, Françoise Lordey, Isabelle Mustapha, Selima Fourati Ben Bouker, Amin Zouari, Raoudha Thierry-Mieg, Nicolas Nef, Serge Arnoult, Christophe Ray, Pierre F. Kherraf, Zine-Eddine Cells Article The genetic landscape of male infertility is highly complex. It is estimated that at least 4000 genes are involved in human spermatogenesis, but only few have so far been extensively studied. In this study, we investigated by whole exome sequencing two cases of idiopathic non-obstructive azoospermia (NOA) due to severe hypospermatogenesis. After variant filtering and prioritizing, we retained for each patient a homozygous loss-of-function (LoF) variant in a testis-specific gene, C1orf185 (c.250C>T; p.Gln84Ter) and CCT6B (c.615-2A>G), respectively. Both variants are rare according to the gnomAD database and absent from our local control cohort (n = 445). To verify the implication of these candidate genes in NOA, we used the CRISPR/Cas9 system to invalidate the mouse orthologs 4930522H14Rik and Cct6b and produced two knockout (KO) mouse lines. Sperm and testis parameters of homozygous KO adult male mice were analyzed and compared with those of wild-type animals. We showed that homozygous KO males were fertile and displayed normal sperm parameters and a functional spermatogenesis. Overall, these results demonstrate that not all genes highly and specifically expressed in the testes are essential for spermatogenesis, and in particular, we conclude that bi-allelic variants of C1orf185 and CCT6B are most likely not to be involved in NOA and male fertility. MDPI 2021-12-30 /pmc/articles/PMC8750304/ /pubmed/35011680 http://dx.doi.org/10.3390/cells11010118 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cazin, Caroline
Neirijnck, Yasmine
Loeuillet, Corinne
Wehrli, Lydia
Kühne, Françoise
Lordey, Isabelle
Mustapha, Selima Fourati Ben
Bouker, Amin
Zouari, Raoudha
Thierry-Mieg, Nicolas
Nef, Serge
Arnoult, Christophe
Ray, Pierre F.
Kherraf, Zine-Eddine
Combined Use of Whole Exome Sequencing and CRISPR/Cas9 to Study the Etiology of Non-Obstructive Azoospermia: Demonstration of the Dispensable Role of the Testis-Specific Genes C1orf185 and CCT6B
title Combined Use of Whole Exome Sequencing and CRISPR/Cas9 to Study the Etiology of Non-Obstructive Azoospermia: Demonstration of the Dispensable Role of the Testis-Specific Genes C1orf185 and CCT6B
title_full Combined Use of Whole Exome Sequencing and CRISPR/Cas9 to Study the Etiology of Non-Obstructive Azoospermia: Demonstration of the Dispensable Role of the Testis-Specific Genes C1orf185 and CCT6B
title_fullStr Combined Use of Whole Exome Sequencing and CRISPR/Cas9 to Study the Etiology of Non-Obstructive Azoospermia: Demonstration of the Dispensable Role of the Testis-Specific Genes C1orf185 and CCT6B
title_full_unstemmed Combined Use of Whole Exome Sequencing and CRISPR/Cas9 to Study the Etiology of Non-Obstructive Azoospermia: Demonstration of the Dispensable Role of the Testis-Specific Genes C1orf185 and CCT6B
title_short Combined Use of Whole Exome Sequencing and CRISPR/Cas9 to Study the Etiology of Non-Obstructive Azoospermia: Demonstration of the Dispensable Role of the Testis-Specific Genes C1orf185 and CCT6B
title_sort combined use of whole exome sequencing and crispr/cas9 to study the etiology of non-obstructive azoospermia: demonstration of the dispensable role of the testis-specific genes c1orf185 and cct6b
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8750304/
https://www.ncbi.nlm.nih.gov/pubmed/35011680
http://dx.doi.org/10.3390/cells11010118
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