Engineering inducible biomolecular assemblies for genome imaging and manipulation in living cells

Qin Peng (), Ziliang Huang, Kun Sun, Yahan Liu, Chi Woo Yoon, Reed E. S. Harrison, Danielle L. Schmitt, Linshan Zhu, Yiqian Wu, Ipek Tasan, Huimin Zhao, Jin Zhang, Sheng Zhong, Shu Chien and Yingxiao Wang ()
Additional contact information
Qin Peng: University of California
Ziliang Huang: University of California
Kun Sun: Institute of Cancer Research, Shenzhen Bay Laboratory
Yahan Liu: University of California
Chi Woo Yoon: University of California
Reed E. S. Harrison: University of California
Danielle L. Schmitt: University of California
Linshan Zhu: University of California
Yiqian Wu: University of California
Ipek Tasan: University of Illinois at Urbana-Champaign
Huimin Zhao: University of Illinois at Urbana-Champaign
Jin Zhang: University of California
Sheng Zhong: University of California
Shu Chien: University of California
Yingxiao Wang: University of California

Nature Communications, 2022, vol. 13, issue 1, 1-15

Abstract: Abstract 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.

Date: 2022
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DOI: 10.1038/s41467-022-35504-x

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