Development of CRISPR-based epigenome editing tools is important for the study and engineering of biological behavior.Here,we describe the design of a reporter system for quantifying the ability of CRISPR epigenome ed...Development of CRISPR-based epigenome editing tools is important for the study and engineering of biological behavior.Here,we describe the design of a reporter system for quantifying the ability of CRISPR epigenome editors to produce a stable gene repression.We characterize the dynamics of durable gene silencing and reactivation,as well as the induced epigenetic changes of this system.We report the creation of single-protein CRISPR constructs bearing combinations of three epigenetic editing domains,termed KAL,that can stably repress the gene expression.This system should allow for the development of novel epigenome editing tools which will be useful in a wide array of biological research and engineering applications.展开更多
基金supported by the Stanford School of Medicine Dean’s Postdoctoral Fellowship.A.E.I.was supported by Stanford ChEM-H Undergraduate Research Fellow and Bio-X REU programs.Y.G.was supported by the National Science Foundation Graduate Research Fellowship Program.L.S.Q.acknowledges support from the Pew Charitable Trusts,the Alfred P.Sloan Foundation,and the Li Ka Shing Foundationby the Li Ka Shing Foundation and partly supported by the National Institutes of Health Common Fund 4D Nucleome Program(U01 DK127405).
文摘Development of CRISPR-based epigenome editing tools is important for the study and engineering of biological behavior.Here,we describe the design of a reporter system for quantifying the ability of CRISPR epigenome editors to produce a stable gene repression.We characterize the dynamics of durable gene silencing and reactivation,as well as the induced epigenetic changes of this system.We report the creation of single-protein CRISPR constructs bearing combinations of three epigenetic editing domains,termed KAL,that can stably repress the gene expression.This system should allow for the development of novel epigenome editing tools which will be useful in a wide array of biological research and engineering applications.
