Kushawah Gopal, Amaral Danielson Baia, Hassan Huzaifa, Gogol Madelaine, Nowotarski Stephanie H, Bazzini Ariel A
Stowers Institute for Medical Research, 1000 E 50th Street, Kansas City, MO 64110, USA.
Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Blvd, Kansas City, KS 66160, USA.
bioRxiv. 2024 Nov 8:2024.11.07.622483. doi: 10.1101/2024.11.07.622483.
The maternal-to-zygotic transition shifts regulatory control from maternal to zygotic messenger RNAs (mRNA) through maternal mRNA degradation. While temporal aspects of maternal mRNA decay are known, spatial mechanisms remain underexplored. Using CRISPR-Cas9 and CRISPR-Cas13d systems, we functionally dissected the contribution of maternal versus zygotic fractions and overcame challenges of studying embryonic lethal genes. We identified differentially distributed maternal mRNAs in specific cells and evidenced the critical role of five maternal mRNAs, and in embryogenesis. Further, we focused on the functionally uncharacterized gene, revealing its essential role in gametogenesis and embryogenesis. acts as a spatio-temporal RNA decay factor regulating mRNA stability and accumulation of its targets in a spatio-temporal manner through 3'UTR recognition during early development. Furthermore, 3'UTR drives its spatio-temporal RNA localization. Our findings provide new insights into spatio-temporal RNA decay mechanisms and highlight dual CRISPR-Cas strategies in studying embryonic development.
母源-合子转变通过母源mRNA降解将调控控制权从母源信使核糖核酸(mRNA)转移至合子mRNA。虽然母源mRNA降解的时间方面已为人所知,但其空间机制仍未得到充分探索。利用CRISPR-Cas9和CRISPR-Cas13d系统,我们从功能上剖析了母源与合子组分的作用,并克服了研究胚胎致死基因的挑战。我们在特定细胞中鉴定出差异分布的母源mRNA,并证明了5种母源mRNA在胚胎发生中的关键作用。此外,我们聚焦于功能未明的基因,揭示了其在配子发生和胚胎发生中的重要作用。在早期发育过程中,通过识别3'非翻译区(3'UTR),作为一种时空RNA衰变因子,以时空方式调节mRNA稳定性及其靶标的积累。此外,3'UTR驱动其时空RNA定位。我们的研究结果为时空RNA衰变机制提供了新见解,并突出了双CRISPR-Cas策略在研究胚胎发育中的作用。