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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 |
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