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Photoactivatable RNA N -Methyladenosine Editing with CRISPR-Cas13

Small. 2020-06-01; 
Jie Zhao, Bing Li, Jianxiong Ma, Weilin Jin, Xinlong Ma
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Codon Optimization … Cloning of orthologues for activity screen and recombinant expression. We synthesized human codon-optimized versions of 15 Cas13a orthologues (Genscript) (Supplementary Table 10) and cloned them into pACYC184 under a pLac promoter … Get A Quote

摘要

RNA has important and diverse biological roles, but the molecular methods to manipulate it spatiotemporally are limited. Here, an engineered photoactivatable RNA N -methyladenosine (m A) editing system with CRISPR-Cas13 is designed to direct specific m A editing. Light-inducible heterodimerizing proteins CIBN and CRY2PHR are fused to catalytically inactive PguCas13 (dCas13) and m A effectors, respectively. This system, referred to as PAMEC, enables the spatiotemporal control of m A editing in response to blue light. Further optimization of this system to create a highly efficient version, known as PAMEC , allows the manipulation of multiple genes robustly and simultaneously. When coupled with an upconversion na... More

關鍵詞

CRISPR-Cas13b, N6-methyladenosine, RNA editing, optogenetic platforms
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