Photoactivatable RNA N⁶‐Methyladenosine Editing with CRISPR‐Cas13

Abstract

RNA has important and diverse biological roles, but the molecular methods to manipulate it spatiotemporally are limited. Here, an engineered photoactivatable RNA N6‐methyladenosine (m6A) editing system with CRISPR‐Cas13 is designed to direct specific m6A editing. Light‐inducible heterodimerizing proteins CIBN and CRY2PHR are fused to catalytically inactive PguCas13 (dCas13) and m6A effectors, respectively. This system, referred to as PAMEC, enables the spatiotemporal control of m6A editing in response to blue light. Further optimization of this system to create a highly efficient version, known as PAMECR, 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.

A Blue Light-Inducible CRISPR-Cas9 System for Inhibiting Progression of Melanoma Cells

“…As a simple and easy-to-obtain light source, blue light is often used for anti-inflammation and sterilization on the skin, without side effects on the human body (Lotan et al, 2019;Zhao J. et al, 2020). In this study, we used blue light as the controller of the CRISPR-Cas9 system.…”

Cross-talk Between Histone and DNA Methylation Mediates Bone Loss in Hind Limb Unloading

“…Cell cultures were carried out as previously described. ( 7,8,23,25 ) Briefly, UMR106 cells and HEK293T cells obtained from Cell Bank of Chinese Academy of Sciences were cultured in Dulbecco's modified Eagle's medium (Gibco, Thermo Fisher Scientific) with 10% fetal bovine serum (Gibco) at 37°C under a humidified atmosphere of 5% CO 2 in air. Primary rat mesenchymal stem cells (MSCs) were isolated by flushing the bone marrow of tibias and femurs.…”

Methyladenosine Modification in RNAs: Classification and Roles in Gastrointestinal Cancers

“…However, the inhibitors of other m 6 A regulators such as METTL3, METTL14, or WTAP have not been systematically developed. Moreover, targeted RNA demethylation or methylation by the engineered dCas13-containing fusion proteins may hold the potential to develop a treatment regimen for GICs ( 174 – 176 ) ( Figure 3 ).…”