CRISPR/Cas9-Mediated Genome Editing Reveals Oosp Family Genes are Dispensable for Female Fertility in Mice.

Ferheen Abbasi,Masashi Mori,Mayo Kodani,Daiji Kiyozumi,Yoshitaka Fujihara,Masahito Ikawa,Chihiro Emori

doi:10.3390/cells9040821

Ferheen Abbasi, Masashi Mori + Show 5 more

Open Access

https://doi.org/10.3390/cells9040821

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Abstract

There are over 200 genes that are predicted to be solely expressed in the oocyte and ovary, and thousands more that have expression patterns in the female reproductive tract. Unfortunately, many of their physiological functions, such as their roles in oogenesis or fertilization, have yet to be elucidated. Previous knockout (KO) mice studies have proven that many of the genes that were once thought to be essential for fertility are dispensable in vivo. Therefore, it is extremely important to confirm the roles of all genes before spending immense time studying them in vitro. To do this, our laboratory analyzes the functions of ovary and oocyte-enriched genes in vivo through generating CRISPR/Cas9 KO mice and examining their fertility. In this study, we have knocked out three Oosp family genes (Oosp1, Oosp2, and Oosp3) that have expression patterns linked to the female reproductive system and found that the triple KO (TKO) mutant mice generated exhibited decreased prolificacy but were not infertile; thus, these genes may potentially be dispensable for fertility. We also generated Cd160 and Egfl6 KO mice and found these genes are individually dispensable for female fertility. KO mice with no phenotypic data are seldom published, but we believe that this information must be shared to prevent unnecessary experimentation by other laboratories.

Highlights

According to a study conducted by the World Health Organization in 2012, 1.9% of women aged20–44 were unable to have their first child [1]
Using a variety of bioinformatic websites, we started by identifying potential target genes that may be important for female reproduction
Once we made a list of all viable candidates, we systematically conducted RT-PCR experimentation to ensure that the gene was enriched in the female reproductive tract

Summary

Introduction

According to a study conducted by the World Health Organization in 2012, 1.9% of women aged20–44 were unable to have their first child [1]. According to a study conducted by the World Health Organization in 2012, 1.9% of women aged. Studies show that women tend to blame themselves for infertility [2], even though half of all cases of infertility are linked to their male counterparts [3]. The genetic bases of female infertility are not fully understood, but it can be linked to ovulation disorders [4], tubal infertility [5], endometriosis [6], and implantation disorders [7], among many others. It has been estimated that 50% of infertility cases are genetic, yet few genes have been shown to cause or to be strongly associated with primary infertility [8], which affects germ cells leading to developmental arrest and cell death. Due to assisted reproductive technologies (ART), many infertile women can successfully have children, depending on their particular cause of infertility

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