Engineering inducible biomolecular assemblies for genome imaging and manipulation in living cells.
Engineering inducible biomolecular assemblies for genome imaging and manipulation in living cells.
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DOI：
10.1038/s41467-022-35504-x
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发表时间：
2022-12-24
影响因子：
16.6
通讯作者：
Wang, Yingxiao
中科院分区：
文献类型：
--
作者：
Peng, Qin;Huang, Ziliang;Sun, Kun;Liu, Yahan;Yoon, Chi Woo;Harrison, Reed E. S.;Schmitt, Danielle L.;Zhu, Linshan;Wu, Yiqian;Tasan, Ipek;Zhao, Huimin;Zhang, Jin;Zhong, Sheng;Chien, Shu;Wang, Yingxiao
基因组结构和组织在细胞生命中起着关键作用。然而，它仍然在很大程度上是未知的基因组位点是如何动态协调，以调节基因表达和决定细胞命运在单细胞水平。我们已经开发了一个诱导系统，它允许同时成像和操纵基因组位点的生物分子组装体（SIMBA）在活细胞中。在SIMBA中，人异染色质蛋白1α（HP 1 α）与mCherry和FRB融合，可以诱导其与FKBP-scFv形成生物分子组装体（BA），并通过核酸酶缺陷型Cas9（dCas 9）或转录因子（TF）引导至特定的基因组位点。携带SunTag串联重复序列。诱导的BA不仅可以使用单个sgRNA在重复或非重复序列处增强靶基因组基因座处的成像信号，而且还可以募集表观遗传调节剂如组蛋白甲基转移酶SUV 39 H1以局部抑制转录。因此，SIMBA原则上可以应用于同时可视化和操纵活细胞中具有可控定时的任何基因组位点。活细胞中的非重复基因座成像仍然具有挑战性。在这里，作者设计了一种诱导系统，由此生物分子组装可以通过核酸酶缺陷型Cas9引导到特定的基因组位点，从而允许同时成像和操纵位点。
Genome architecture and organization play critical roles in cell life. However, it remains largely unknown how genomic loci are dynamically coordinated to regulate gene expression and determine cell fate at the single cell level. We have developed an inducible system which allows Simultaneous Imaging and Manipulation of genomic loci by Biomolecular Assemblies (SIMBA) in living cells. In SIMBA, the human heterochromatin protein 1α (HP1α) is fused to mCherry and FRB, which can be induced to form biomolecular assemblies (BAs) with FKBP-scFv, guided to specific genomic loci by a nuclease-defective Cas9 (dCas9) or a transcriptional factor (TF) carrying tandem repeats of SunTag. The induced BAs can not only enhance the imaging signals at target genomic loci using a single sgRNA, either at repetitive or non-repetitive sequences, but also recruit epigenetic modulators such as histone methyltransferase SUV39H1 to locally repress transcription. As such, SIMBA can be applied to simultaneously visualize and manipulate, in principle, any genomic locus with controllable timing in living cells. Imaging non-repetitive loci in living cells remains challenging. Here, the authors engineered an inducible system whereby biomolecular assemblies can be guided to specific genomic loci by a nuclease-defective Cas9, allowing the simultaneous imaging and manipulation of the loci.
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影响因子：
16.6
作者：
Braun SMG;Kirkland JG;Chory EJ;Husmann D;Calarco JP;Crabtree GR
通讯作者：
Crabtree GR
影响因子：
7
作者：
Chaudhary N;Nho SH;Cho H;Gantumur N;Ra JS;Myung K;Kim H
通讯作者：
Kim H
影响因子：
56.9
作者：
Frey, Steffen;Richter, Ralf P.;Goerlich, Dirk
通讯作者：
Goerlich, Dirk
DOI：
10.1126/science.aao3136
发表时间：
2018-03-02
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gu B;Swigut T;Spencley A;Bauer MR;Chung M;Meyer T;Wysocka J
通讯作者：
Wysocka J
影响因子：
16
作者：
Gao Y;Han M;Shang S;Wang H;Qi LS
通讯作者：
Qi LS