TY - JOUR
T1 - Engineering inducible biomolecular assemblies for genome imaging and manipulation in living cells
AU - Peng, Qin
AU - Huang, Ziliang
AU - Sun, Kun
AU - Liu, Yahan
AU - Yoon, Chi Woo
AU - Harrison, Reed E.S.
AU - Schmitt, Danielle L.
AU - Zhu, Linshan
AU - Wu, Yiqian
AU - Tasan, Ipek
AU - Zhao, Huimin
AU - Zhang, Jin
AU - Zhong, Sheng
AU - Chien, Shu
AU - Wang, Yingxiao
N1 - We thank Professors Bing Ren and Wei Wang for the discussion of the experimental design. This publication includes data generated at the UC San Diego IGM Genomics Center utilizing an Illumina NovaSeq 6000 that was purchased with funding from a National Institutes of Health SIG grant (#S10 OD026929). We would like to thank the UCSD Nikon Imaging Center for assistance with imaging. This work was supported in part by grants from NIH R01 EB029122, R01 HD107206, R35 GM140929 (Y.W.), R01 HL121365 (S.C. and Y.W.), R35 CA197622 (J.Z.), U54DK107965 (H.Z.), and San Diego IRACDA support to D.L.S. (GM068524).
PY - 2022/12
Y1 - 2022/12
N2 - 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.
AB - 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.
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U2 - 10.1038/s41467-022-35504-x
DO - 10.1038/s41467-022-35504-x
M3 - Article
C2 - 36566209
AN - SCOPUS:85144638485
SN - 2041-1723
VL - 13
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 7933
ER -