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Photoactivatable RNA N 6 ‐Methyladenosine Editing with CRISPR‐Cas13
Author(s) -
Zhao Jie,
Li Bing,
Ma Jianxiong,
Jin Weilin,
Ma Xinlong
Publication year - 2020
Publication title -
small
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.785
H-Index - 236
eISSN - 1613-6829
pISSN - 1613-6810
DOI - 10.1002/smll.201907301
Subject(s) - optogenetics , crispr , rna , genome editing , guide rna , computational biology , computer science , nanotechnology , biology , gene , materials science , genetics , neuroscience
RNA has important and diverse biological roles, but the molecular methods to manipulate it spatiotemporally are limited. Here, an engineered photoactivatable RNA N 6 ‐methyladenosine (m 6 A) editing system with CRISPR‐Cas13 is designed to direct specific m 6 A editing. Light‐inducible heterodimerizing proteins CIBN and CRY2PHR are fused to catalytically inactive PguCas13 (dCas13) and m 6 A effectors, respectively. This system, referred to as PAMEC, enables the spatiotemporal control of m 6 A editing in response to blue light. Further optimization of this system to create a highly efficient version, known as PAMEC R , allows the manipulation of multiple genes robustly and simultaneously. When coupled with an upconversion nanoparticle film, the optogenetic operation window is extended from the visible range to tissue‐penetrable near‐infrared wavelengths, which offers an appealing avenue to remotely control RNA editing. These results show that PAMEC is a promising optogenetic platform for flexible and efficient targeting of RNA, with broad applicability for epitranscriptome engineering, imaging, and future therapeutic development.