CRISPR-assisted detection of RNA-protein interactions in living cells
CRISPR-assisted detection of RNA-protein interactions in living cells
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CRISPR 辅助检测活细胞中 RNA-蛋白质相互作用
DOI：
10.1038/s41592-020-0866-0
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发表时间：
2020-06-22
期刊：
影响因子：
48
通讯作者：
Yan, Jian
中科院分区：
文献类型：
--
作者：
Yi, Wenkai;Li, Jingyu;Yan, Jian
CARPID利用CRISPR技术将生物素连接酶定位到特定的lncRNA上，从而实现近端标记，从而查询活细胞中RNA-蛋白质相互作用。我们开发了CRISPR辅助的RNA-蛋白质相互作用检测方法（CARPID），该方法利用基于CRISPR-CasRx的RNA靶向和近端标记来识别天然细胞背景下特定长非编码RNA（lncRNA）的结合蛋白。我们将CARPID应用于核lncRNA XIST，它捕获了一系列已知的相互作用蛋白和多种以前未表征的结合蛋白。我们将CARPID推广到探索lncRNAs DANCR和MALAT1的结合物，揭示了该方法在识别RNA结合蛋白方面的广泛适用性。
CARPID uses CRISPR technology to navigate biotin ligase to specific lncRNAs, which allows proximal labeling and thus the querying of RNA-protein interactions in living cells.We have developed CRISPR-assisted RNA-protein interaction detection method (CARPID), which leverages CRISPR-CasRx-based RNA targeting and proximity labeling to identify binding proteins of specific long non-coding RNAs (lncRNAs) in the native cellular context. We applied CARPID to the nuclear lncRNA XIST, and it captured a list of known interacting proteins and multiple previously uncharacterized binding proteins. We generalized CARPID to explore binders of the lncRNAs DANCR and MALAT1, revealing the method's wide applicability in identifying RNA-binding proteins.