期刊
NUCLEIC ACIDS RESEARCH
卷 45, 期 18, 页码 10800-10810出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkx675
关键词
-
资金
- National Institutes of Health (NIH) [R01HL124285, R01GM110251, K25HL121295, R01CA197139, R01MH109907, P01HD084387, UM1HG009408]
- NIH [R01HL124285, R01GM110251, K25HL121295, R01CA197139, R01MH109907, P01HD084387, UM1HG009408]
Many studies using reporter assays have demonstrated that 3' untranslated regions (3'-UTRs) regulate gene expression by controlling mRNA stability and translation. Due to intrinsic limitations of heterologous reporter assays, we sought to develop a gene editing approach to investigate the regulatory activity of 3'-UTRs in their native context. We initially used dual-CRISPR (clustered, regularly interspaced, short palindromic repeats)-Cas9 targeting to delete DNA regions corresponding to nine chemokine 3' UTRs that destabilized mRNAin a reporter assay. Targeting six chemokine 3'-UTRs increased chemokine mRNAlevels as expected. However, targetingCXCL1, CXCL6 and CXCL8 3'-UTRs unexpectedly led to substantial mRNA decreases. Metabolic labeling assays showed that targeting these three 3'-UTRs increased mRNA stability, as predicted by the reporter assay, while also markedly decreasing transcription, demonstrating an unexpected role for 3'-UTR sequences in transcriptional regulation. We further show that CRISPR-Cas9 targeting of specific ' UTR elements can be used for modulating gene expression and for highly parallel localization of active 3'-UTR elements in the native context. Our work demonstrates the duality and complexity of 3'-UTR sequences in regulation of gene expression and provides a useful approach for modulating gene expression and for functional annotation of 3'-UTRs in the native context.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据