期刊
NATURE METHODS
卷 16, 期 7, 页码 633-+出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/s41592-019-0436-5
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资金
- New York Stem Cell Foundation
- Alfred P. Sloan Foundation
- NIH Director's New Innovator Award from the National Institute of Mental Health
- 4D Nucleome Common Fund grant [1U01HL12999801]
- NSF-NIGMS [1562665]
- National Science Foundation Graduate Research Fellowship [DGE-1321851]
- [1DP2MH11024701]
- Division Of Mathematical Sciences
- Direct For Mathematical & Physical Scien [1562665] Funding Source: National Science Foundation
Mammalian genomes are folded into tens of thousands of long-range looping interactions. The cause-and-effect relationship between looping and genome function is poorly understood, and the extent to which loops are dynamic on short time scales remains an unanswered question. Here, we engineer a new class of synthetic architectural proteins for directed rearrangement of the three-dimensional genome using blue light. We target our light-activated-dynamic-looping (LADL) system to two genomic anchors with CRISPR guide RNAs and induce their spatial colocalization via light-induced heterodimerization of cryptochrome 2 and a dCas9-CIBN fusion protein. We apply LADL to redirect a stretch enhancer (SE) away from its endogenous Klf4 target gene and to the Zfp462 promoter. Using single-molecule RNA-FISH, we demonstrate that de novo formation of the Zfp462-SE loop correlates with a modest increase in Zfp462 expression. LADL facilitates colocalization of genomic loci without exogenous chemical cofactors and will enable future efforts to engineer reversible and oscillatory loops on short time scales.
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