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Engineering an adenine base editor in human embryonic stem cells with minimal DNA and RNA off-target activities

Mol Ther Nucleic Acids. 2022-08; 
Zhenwu Zhang, Wanyu Tao, Shisheng Huang, Wenjun Sun, Yue Wang, Wen Jiang, Xingxu Huang, Chao-Po Lin
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GenParts™ DNA Fragments … To construct base-editor expression plasmids, TadA8e were synthesized by GenScript and … , the TRE3Gs fragment was synthesized by GenScript and cloned into a piggyBac plasmid … Get A Quote

摘要

Genome editing in pluripotent stem cells (PSCs) using CRISPR technology holds great promise for therapeutic applications. Yet, it has been reported that Cas9-mediated cleavage could cause large deletions or rearrangements of DNA, and the selection of edited PSCs could acquire p53 mutations. Adenine base editors (ABEs) do not introduce DNA double-strand breaks and thus have been proposed as alternatives to circumvent those problems, but their off-target effects still limit their applications. Here, we tested different combinations of off-target reduction methods to further diminish off-target effects of ABEs without compromising their on-target editing efficiencies. We subsequently chose the best editor, CE-8e-d... More

关键词

CRISPR, MT: RNA/DNA Editing, adenine base editor, human embryonic stem cell, off-target effect