Regulation of gene expression in the bovine blastocyst by CSF2 is disrupted by CRISPR/Cas9-mediated deletion of CSF2RA.

Regulation of gene expression in the bovine blastocyst by CSF2 is disrupted by CRISPR/Cas9-mediated deletion of CSF2RA. Biol Reprod. 2021 Feb 02;: Authors: Xiao Y, Uh K, Negrón-Pérez VM, Haines H, Lee K, Hansen PJ Abstract Colony stimulating factor 2 (CSF2) functions in the reproductive tract to modulate function of the preimplantation embryo. The β subunit of the CSF2 receptor (CSF2RB) is not expressed in the embryo and signal transduction is therefore different than for myeloid cells where the receptor is composed of α (CSF2RA) and β subunits. Here, we produced embryos in which exons 5 and 6 of CSF2RA were disrupted using the CRISPR/Cas 9 system to test whether CSF2RA signaling was essential for actions of CSF2 in the bovine embryo. Wildtype and CSF2RA knockout embryos were treated with 10 ng/mL CSF2 or vehicle at day 5 of development. Blastocysts were harvested at day 8 to determine transcript abundance of 90 genes by real time PCR. Responses in female blastocysts were examined separately from male blastocysts because actions of CSF2 are sex-dependent. For wildtype embryos, CSF2 altered expression of 10 genes in females and 20 in males. Only three genes were affected by CSF2 in a similar manner for both sexes. Disruption of CSF2RA prevented the effect of CSF2 on expression for 9 of 10 CSF2-regulated genes in females and 19 of 20 genes in males. Results confirm the importance of CSF2RA for regulation of gene expression by CSF...
Source: Biology of Reproduction - Category: Reproduction Medicine Authors: Tags: Biol Reprod Source Type: research