小柯机器人

美国哈佛大学刘如谦（David R. Liu）研究小组利用噬菌体辅助进化，产生了具有Cas结构域兼容性与活性提升的腺嘌呤碱基编辑器（ABE）。2020年3月17日，《自然—生物技术》杂志在线发表了这一最新研究成果。

Title: Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility and activity

Author: Michelle F. Richter, Kevin T. Zhao, Elliot Eton, Audrone Lapinaite, Gregory A. Newby, Benjamin W. Thuronyi, Christopher Wilson, Luke W. Koblan, Jing Zeng, Daniel E. Bauer, Jennifer A. Doudna, David R. Liu

Issue&Volume: 2020-03-16

Abstract: Applications of adenine base editors (ABEs) have been constrained by the limited compatibility of the deoxyadenosine deaminase component with Cas homologs other than SpCas9. We evolved the deaminase component of ABE7.10 using phage-assisted non-continuous and continuous evolution (PANCE and PACE), which resulted in ABE8e. ABE8e contains eight additional mutations that increase activity (kapp) 590-fold compared with that of ABE7.10. ABE8e offers substantially improved editing efficiencies when paired with a variety of Cas9 or Cas12 homologs. ABE8e is more processive than ABE7.10, which could benefit screening, disruption of regulatory regions and multiplex base editing applications. A modest increase in Cas9-dependent and -independent DNA off-target editing, and in transcriptome-wide RNA off-target editing can be ameliorated by the introduction of an additional mutation in the TadA-8e domain. Finally, we show that ABE8e can efficiently install natural mutations that upregulate fetal hemoglobin expression in the BCL11A enhancer or in the the HBG promoter in human cells, targets that were poorly edited with ABE7.10. ABE8e augments the effectiveness and applicability of adenine base editing.

DOI: 10.1038/s41587-020-0453-z

Source: https://www.nature.com/articles/s41587-020-0453-z

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：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex