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

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.

Genetic incompatibility of the reproductive partners: an evolutionary perspective on infertility

“… Rohlfs et al (2010) also demonstrated in humans that the zona pellucida (glycoprotein layer surrounding the oocyte) gene ZP3 coevolves with its putative binding partner, ZP3R , in sperm ( Table I ). Furthermore, recent genome editing studies have revealed several other sperm- and oocyte-specific genes that play an important role in gamete interaction ( Abbasi et al , 2020 ; Fujihara et al , 2020 ; Lamas-Toranzo et al , 2020 ; Noda et al , 2020 ). Although the binding partners of these genes remain to be demonstrated in future studies, all of them have a potential to increase our understanding of the molecular mechanisms of gamete incompatibility.…”

Establishment and Phenotypic Analysis of the Novel Gaucher Disease Mouse Model With the Partially Humanized Gba1 Gene and F213I Mutation

“…Crispr/Cas9 system was used to reduce the interference of the endogenous GCase activity, as we performed previously ( Abbasi et al, 2020 ). Briefly, HEK293 cells were infected with lentivirus expressing CAS9 protein and sgRNA of GBA1.…”

Where are all the egg genes?

“…In parallel with characterization of genetic mutants, Whole Genome Sequencing (WGS) can be employed to identify the causative mutation. Another approach being undertaken is to use CRISPR-Cas9 to generate knockouts of all testes expressed genes as is being undertaken by several labs to ascertain which knockouts impact fertilization (Miyata et al, 2016;Abbasi et al, 2020). A similar approach could be taken to knock out all oocyte expressed genes.…”