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Compact RNA editors with small Cas13 proteins

Nat Biotechnol. 2021-08; 
Soumya Kannan, Han Altae-Tran, Xin Jin, Victoria J Madigan, Rachel Oshiro, Kira S Makarova, Eugene V Koonin, Feng Zhang
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Codon Optimization … and cloned into the BamHI site of pACYC184 by GenScript. To clone bacterial expression plasmids for the PFS screen, Cas13bt protein CDSs were human codon optimized using GeneArt GeneOptimizer (Thermo Fisher) and synthesized by GenScript into a pcDNA3.1(+) … Get A Quote

摘要

CRISPR-Cas13 systems have been developed for precise RNA editing, and can potentially be used therapeutically when temporary changes are desirable or when DNA editing is challenging. We have identified and characterized an ultrasmall family of Cas13b proteins-Cas13bt-that can mediate mammalian transcript knockdown. We have engineered compact variants of REPAIR and RESCUE RNA editors by functionalizing Cas13bt with adenosine and cytosine deaminase domains, and demonstrated packaging of the editors within a single adeno-associated virus.

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