Gene-specific quantification of nascent transcription following targeted degradation of endogenous proteins in cultured cells.
Gene-specific quantification of nascent transcription following targeted degradation of endogenous proteins in cultured cells.
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DOI：
10.1016/j.xpro.2021.101000
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发表时间：
2021-12-17
期刊：
影响因子：
--
通讯作者：
Tansey WP
中科院分区：
文献类型：
--
作者：
Guarnaccia AD;Weissmiller AM;Tansey WP
全基因组核连续运行是确定转录因子降解等扰动对转录模式影响的有效方法。但研究人员通常感兴趣的是监测特定基因组的转录效应，而不是整个基因组。在这里，我们描述了一种方法，该方法将基因组工程与标记内源蛋白进行降解结合起来，通过简化的核连续测定来产生有关因子耗尽引起的初级转录变化的基因特异性信息。有关该协议的使用和执行的完整详细信息，请参阅。降解蛋白质和分析基因特异性转录效应的策略 CRISPR 标签能够快速降解培养细胞中的靶蛋白 基因特异性核连载 (NRO) 监测感兴趣转录本的变化 简化从细胞系工程到 NRO 数据分析的工作流程 全基因组核连连是确定转录因子降解等扰动对转录模式影响的有效方法。但研究人员通常感兴趣的是监测特定基因组的转录效应，而不是整个基因组。在这里，我们描述了一种方法，该方法将基因组工程与标记内源蛋白进行降解结合起来，通过简化的核连续测定来产生有关因子耗尽引起的初级转录变化的基因特异性信息。
Genome-wide nuclear run-ons are a powerful way to determine the impact of a perturbation such as transcription factor degradation on transcriptional patterns. But often investigators are interested in monitoring transcriptional effects at specific sets of genes, rather than the entire genome. Here we describe an approach that couples genome engineering to tag endogenous proteins for degradation with a streamlined nuclear run-on assay to yield gene-specific information on primary transcriptional changes elicited by factor depletion. For complete details on the use and execution of this protocol, please refer to. Strategy for degrading a protein and analyzing gene-specific transcriptional effects CRISPR tagging enables rapid degradation of a target protein in cultured cells Gene-specific nuclear run-on (NRO) monitors changes in transcripts of interest Streamlined workflow from cell line engineering through to NRO data analysis Genome-wide nuclear run-ons are a powerful way to determine the impact of a perturbation such as transcription factor degradation on transcriptional patterns. But often investigators are interested in monitoring transcriptional effects at specific sets of genes, rather than the entire genome. Here we describe an approach that couples genome engineering to tag endogenous proteins for degradation with a streamlined nuclear run-on assay to yield gene-specific information on primary transcriptional changes elicited by factor depletion.
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影响因子：
14.8
作者：
Nabet B;Roberts JM;Buckley DL;Paulk J;Dastjerdi S;Yang A;Leggett AL;Erb MA;Lawlor MA;Souza A;Scott TG;Vittori S;Perry JA;Qi J;Winter GE;Wong KK;Gray NS;Bradner JE
通讯作者：
Bradner JE
影响因子：
16.6
作者：
Weissmiller, April M.;Wang, Jing;Tansey, William P.
通讯作者：
Tansey, William P.
影响因子：
14.9
作者：
Concordet JP;Haeussler M
通讯作者：
Haeussler M
影响因子：
7.7
作者：
Popay TM;Wang J;Adams CM;Howard GC;Codreanu SG;Sherrod SD;McLean JA;Thomas LR;Lorey SL;Machida YJ;Weissmiller AM;Eischen CM;Liu Q;Tansey WP
通讯作者：
Tansey WP
DOI：
10.1073/pnas.1910391116
发表时间：
2019-12-10
影响因子：
11.1
作者：
Thomas, Lance R.;Adams, Clare M.;Tansey, William P.
通讯作者：
Tansey, William P.