Simple and versatile imaging of genomic loci in live mammalian cells and early pre-implantation embryos using CAS-LiveFISH.
Simple and versatile imaging of genomic loci in live mammalian cells and early pre-implantation embryos using CAS-LiveFISH.
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DOI：
10.1038/s41598-021-91787-y
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发表时间：
2021-06-09
影响因子：
4.6
通讯作者：
Pertsinidis A
中科院分区：
文献类型：
--
作者：
Geng Y;Pertsinidis A
Visualizing the 4D genome in live cells is essential for understanding its regulation. Programmable DNA-binding probes, such as fluorescent clustered regularly interspaced short palindromic repeats (CRISPR) and transcription activator-like effector (TALE) proteins have recently emerged as powerful tools for imaging specific genomic loci in live cells. However, many such systems rely on genetically-encoded components, often requiring multiple constructs that each must be separately optimized, thus limiting their use. Here we develop efficient and versatile systems, based on in vitro transcribed single-guide-RNAs (sgRNAs) and fluorescently-tagged recombinant, catalytically-inactivated Cas9 (dCas9) proteins. Controlled cell delivery of pre-assembled dCas9-sgRNA ribonucleoprotein (RNP) complexes enables robust genomic imaging in live cells and in early mouse embryos. We further demonstrate multiplex tagging of up to 3 genes, tracking detailed movements of chromatin segments and imaging spatial relationships between a distal enhancer and a target gene, with nanometer resolution in live cells. This simple and effective approach should facilitate visualizing chromatin dynamics and nuclear architecture in various living systems.
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影响因子：
16.6
作者：
Fu Y;Rocha PP;Luo VM;Raviram R;Deng Y;Mazzoni EO;Skok JA
通讯作者：
Skok JA
DOI：
10.1126/science.aaq1392
发表时间：
2018-04-20
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Liu TL;Upadhyayula S;Milkie DE;Singh V;Wang K;Swinburne IA;Mosaliganti KR;Collins ZM;Hiscock TW;Shea J;Kohrman AQ;Medwig TN;Dambournet D;Forster R;Cunniff B;Ruan Y;Yashiro H;Scholpp S;Meyerowitz EM;Hockemeyer D;Drubin DG;Martin BL;Matus DQ;Koyama M;Megason SG;Kirchhausen T;Betzig E
通讯作者：
Betzig E
影响因子：
--
作者：
Churchman, L Stirling;Spudich, James A
通讯作者：
Spudich, James A
DOI：
10.1073/pnas.1420024112
发表时间：
2015-03-10
影响因子：
11.1
作者：
Ma, Hanhui;Naseri, Ardalan;Pederson, Thoru
通讯作者：
Pederson, Thoru
影响因子：
6.4
作者：
Dooley KE;Warburton A;McBride AA
通讯作者：
McBride AA