据了解，基础的腺嘌呤碱基编辑器（例如，ABE7.10）能够编辑A T到G C的点突变，但在修饰原代人类细胞中的基因座时，编辑效率可能很低。 附：英文原文 Title: Directed evolution of adenine base editors with increased activity and therapeutic application
Author: Nicole M. Gaudelli, Dieter K. Lam, Holly A. Rees, Noris M. Sol-Esteves, Luis A. Barrera, David A. Born, Aaron Edwards, Jason M. Gehrke, Seung-Joo Lee, Alexander J. Liquori, Ryan Murray, Michael S. Packer, Conrad Rinaldi, Ian M. Slaymaker, Jonathan Yen, Lauren E. Young, Giuseppe Ciaramella
Issue Volume: -04-13
Abstract: The foundational adenine base editors (for example, ABE7.10) enable programmable AT to GC point mutations but editing efficiencies can be low at challenging loci in primary human cells. Here we further evolve ABE7.10 using a library of adenosine deaminase variants to create ABE8s. At NGG protospacer adjacent motif (PAM) sites, ABE8s result in ~1.5 higher editing at protospacer positions A5 A7 and ~3.2 higher editing at positions A3 A4 and A8 A10 compared with ABE7.10. Non-NGG PAM variants have a ~4.2-fold overall higher on-target editing efficiency than ABE7.10. In human CD34+ cells, ABE8 can recreate a natural allele at the promoter of the -globin genes HBG1 and HBG2 with up to 60% efficiency, causing persistence of fetal hemoglobin. In primary human T cells, ABE8s achieve 98 99% target modification, which is maintained when multiplexed across three loci. Delivered as messenger RNA, ABE8s induce no significant levels of single guide RNA (sgRNA)-independent off-target adenine deamination in genomic DNA and very low levels of adenine deamination in cellular mRNA.
DOI: 10.1038/s41587-020-0491-6
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期刊信息
Nature Biotechnology：《自然 生物技术》，创刊于1996年。隶属于施普林格 自然出版集团，最新IF：31.864
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