核糖核酸
DNA修复
生物
细胞生物学
信使核糖核酸
基因敲除
DNA
分子生物学
报告基因
基因
突变体
基因表达
遗传学
作者
Lisa Tschage,Eric Kowarz,Rolf Marschalek
出处
期刊:The CRISPR journal
[Mary Ann Liebert]
日期:2023-06-01
卷期号:6 (3): 289-301
标识
DOI:10.1089/crispr.2022.0105
摘要
"RNA-templated/directed DNA repair" is a biological mechanism that has been experimentally demonstrated in bacteria, yeast, and mammalian cells. Recent study has shown that small noncoding RNAs (DDRNAs) and/or newly RNAPII transcribed RNAs (dilncRNAs) are orchestrating the initial steps of double-strand break (DSB) repair. In this study, we demonstrate that also pre-mRNA could be used as direct or indirect substrate for DSB repair. Our test system is based on (1) a stably integrated mutant reporter gene that produces constitutively a nonspliceable pre-mRNA, (2) a transiently expressed sgRNA-guided dCas13b::ADAR fusion protein to specifically RNA edit the nonspliceable pre-mRNA, and (3) transiently expressed I-SceI to create a DSB situation to study the effect of spliceable pre-mRNA on DNA repair. Based on our data, the RNA-edited pre-mRNA was used in cis for the DSB repair process, thereby converting the genomically encoded mutant reporter gene into an active reporter gene. Overexpression and knockdown of several cellular proteins were performed to delineate their role in this novel "RNA-mediated end joining" pathway.
科研通智能强力驱动
Strongly Powered by AbleSci AI