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bioRxiv · 10.1101/614859

Targeted mRNA demethylation using an engineered dCas13b-ALKBH5 fusion protein

Li, J.·Chen, Z.·Chen, F.·Ling, Y.·Peng, Y.·Luo, N.·Wang, H.
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Abstract

Studies on biological functions of N6-methyladenosine (m6A) modification in mRNA have sprung up in recent years. Here we construct and characterize a CRISPR-Cas13b-based tool for the first time that targeted m6A methylation of mRNA by fusing the catalytically dead Type VI-B Cas13 enzyme from Prevotella sp.P5-125 (dPspCas13b) with the m6A demethylase ALKBH5, which is named as dm6ACRISPR. Subsequently, such system is shown to specific demethylase the m6A of target mRNA such as CYB5A to increase its mRNA stability. In addition, the dm6ACRISPR system appeared to afford efficient demethylation of the target genes with tenuous off-target effects. Together, we provide a programmable and in vivo manipulation tool to study mRNA modification and its potential biological functions of specific gene.

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Li, J., Chen, Z., Chen, F., Ling, Y., Peng, Y., Luo, N., Wang, H.. 2019-04-20. Targeted mRNA demethylation using an engineered dCas13b-ALKBH5 fusion protein. https://doi.org/10.1101/614859

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