Site‐Specific m<sup>6</sup>A Erasing via Conditionally Stabilized CRISPR‐Cas13b Editor
Ying Xu, Yufan Wang, Fu‐Sen Liang
Abstract
Abstract N6‐methyladenosine (m 6 A) on RNAs plays an important role in regulating various biological processes and CRIPSR technology has been employed for programmable m 6 A editing. However, the bulky size of CRISPR protein and constitutively expressed CRISPR/RNA editing enzymes can interfere with the native function of target RNAs and cells. Herein, we reported a conditional m 6 A editing platform (FKBP*‐dCas13b‐ALK) based on a ligand stabilized dCas13 editor. The inducible expression of this m 6 A editing system was achieved by adding or removing the Shield‐1 molecule. We further demonstrated that the targeted recruitment of dCas13b‐m 6 A eraser fusion protein and site‐specific m 6 A erasing were achieved under the control of Shield‐1. Moreover, the release and degradation of dCas13b fusion protein occurred faster than the restoration of m 6 A on the target RNAs after Shield‐1 removal, which provides an ideal opportunity to study the m 6 A function with minimal steric interference from bulky dCas13b fusion protein.