Photoactivatable RNA N⁶ -Methyladenosine Editing with CRISPR-Cas13

RNA has important and diverse biological roles, but the molecular methods to manipulate it spatiotemporally are limited. Here, an engineered photoactivatable RNA N⁶ -methyladenosine (m⁶ A) editing system with CRISPR-Cas13 is designed to direct specific m⁶ A editing. Light-inducible heterodimerizing proteins CIBN and CRY2PHR are fused to catalytically inactive PguCas13 (dCas13) and m⁶ A effectors, respectively. This system, referred to as PAMEC, enables the spatiotemporal control of m⁶ 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.