Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system.
Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system.
复制标题
DOI：
10.1016/j.cell.2013.12.001
复制
发表时间：
2013-12-19
期刊：
影响因子：
64.5
通讯作者：
Huang B
中科院分区：
文献类型：
--
作者：
Chen B;Gilbert LA;Cimini BA;Schnitzbauer J;Zhang W;Li GW;Park J;Blackburn EH;Weissman JS;Qi LS;Huang B
The spatiotemporal organization and dynamics of chromatin play critical roles in regulating genome function. However, visualizing specific, endogenous genomic loci remains challenging in living cells. Here, we demonstrate such an imaging technique by repurposing the bacterial CRISPR/Cas system. Using an EGFP-tagged endonuclease-deficient Cas9 protein and a structurally optimized small guide (sg) RNA, we show robust imaging of repetitive elements in telomeres and coding genes in living cells. Furthermore, an array of sgRNAs tiling along the target locus enables the visualization of non-repetitive genomic sequences. Using this method, we have studied telomere dynamics during elongation or disruption, the subnuclear localization of the MUC4 loci, the cohesion of replicated MUC4 loci on sister chromatids, and their dynamic behaviors during mitosis. This CRISPR imaging tool has potential to significantly improve the capacity to study the conformation and dynamics of native chromosomes in living human cells.
登录
查看更多内容
影响因子：
64.5
作者：
Misteli T
通讯作者：
Misteli T
影响因子：
17.3
作者：
Gaj, Thomas;Gersbach, Charles A.;Barbas, Carlos F., III
通讯作者：
Barbas, Carlos F., III
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
30.8
作者：
Kim, SH;Kaminker, P;Campisi, J
通讯作者：
Campisi, J
通过反式编码的小 RNA 和宿主因子 RNase III 进行 CRISPR RNA 成熟。
影响因子：
64.8
作者：
Deltcheva, Elitza;Chylinski, Krzysztof;Sharma, Cynthia M.;Gonzales, Karine;Chao, Yanjie;Pirzada, Zaid A.;Eckert, Maria R.;Vogel, Joerg;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle