Inhibition of unintended mutations in gene editing
AssigneeShanghaitech University
InventorsJia Chen, Bei Yang, Li Yang, Xingxu Huang, Lijie Wang
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Abstract
Provided are fusion proteins and related molecules useful for conducting base editing with reduced or no off-target mutations. The fusion proteins may include a first fragment comprising a nucleobase deaminase or a catalytic domain thereof, a second fragment comprising a nucleobase deaminase inhibitory domain, and a protease cleavage site between the first fragment and the second fragment. Also provided are improved prime editing systems, including prime editing guide RNA with improved stability.
Claims14
A fusion protein comprising:
- a first fragment comprising a cytidine deaminase or a catalytic domain thereof,
- a second fragment comprising an inhibitor to the cytidine deaminase or catalytic domain thereof, wherein said inhibitor is an inhibitory domain from the cytidine deaminase or from a second cytidine deaminase, and
- a protease cleavage site between the first fragment and the second fragment, wherein protease cleavage site is a protease cleavage site of a protease selected from the group consisting of TEV protease, TuMV protease, PPV protease, PVY protease, ZIKV protease and WNV protease, or is a self-cleavage site.
A fusion protein comprising a first fragment comprising:
- a cytidine deaminase or a catalytic domain thereof,
- an inhibitor to the cytidine deaminase or catalytic domain thereof, wherein said inhibitor is an inhibitory domain from a cytidine deaminase,
- a first RNA recognition peptide, and
- a TEV protease cleavage site between the cytidine deaminase or a catalytic domain thereof and the cytidine deaminase inhibitor.
A method for editing a nucleic acid, comprising contact the nucleic acid with a fusion protein comprising a first fragment comprising a cytidine deaminase or a catalytic domain thereof, cleavably fused to a second fragment comprising an inhibitor to the cytidine deaminase or catalytic domain thereof, wherein said inhibitor is an inhibitor domain from a cytidine deaminase, and wherein the editing is conducted with a protease that cleaves the second fragment from the first fragment thereby activating the nucleobase deaminase or catalytic domain thereof.
Dependencies are inferred from the claim text.