Programmable protein expression using a genetically encoded m<sup>6</sup>A sensor.

Marayati, Bahjat F; Thompson, Matthew G; Holley, Christopher L; Horner, Stacy M; Meyer, Kate D · Nat Biotechnol · 2024

basic_science · Level V

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Abstract

The N<sup>6</sup>-methyladenosine (m<sup>6</sup>A) modification is found in thousands of cellular mRNAs and is a critical regulator of gene expression and cellular physiology. m<sup>6</sup>A dysregulation contributes to several human diseases, and the m<sup>6</sup>A methyltransferase machinery has emerged as a promising therapeutic target. However, current methods for studying m<sup>6</sup>A require RNA isolation and do not provide a real-time readout of mRNA methylation in living cells. Here we present a genetically encoded m<sup>6</sup>A sensor (GEMS) technology, which couples a fluorescent signal with cellular mRNA methylation. GEMS detects changes in m<sup>6</sup>A caused by pharmacological inhibition of the m<sup>6</sup>A methyltransferase, giving it potential utility for drug discovery efforts. Additionally, GEMS can be programmed to achieve m<sup>6</sup>A-dependent delivery of custom protein payloads in cells. Thus, GEMS is a versatile platform for m<sup>6</sup>A sensing that provides both a simple readout for m<sup>6</sup>A methylation and a system for m<sup>6</sup>A-coupled protein expression.

Medical subject headings