Jeong You Kyeong, Lee SeokHoon, Hwang Gue-Ho, Hong Sung-Ah, Park Se-Eun, Kim Jin-Soo, Woo Jae-Sung, Bae Sangsu
Department of Chemistry and Research Institute for Convergence of Basic Sciences, Hanyang University, Seoul, South Korea.
Center for Genome Engineering, Institute for Basic Science, Daejeon, South Korea.
Nat Biotechnol. 2021 Nov;39(11):1426-1433. doi: 10.1038/s41587-021-00943-2. Epub 2021 Jul 1.
Adenine base editors (ABEs) catalyze specific A-to-G conversions at genomic sites of interest. However, ABEs also induce cytosine deamination at the target site. To reduce the cytosine editing activity, we engineered a commonly used adenosine deaminase, TadA7.10, and found that ABE7.10 with a D108Q mutation in TadA7.10 exhibited tenfold reduced cytosine deamination activity. The D108Q mutation also reduces cytosine deamination activity in two recently developed high-activity versions of ABE, ABE8e and ABE8s, and is compatible with V106W, a mutation that reduces off-target RNA editing. ABE7.10 containing a P48R mutation displayed increased cytosine deamination activity and a substantially reduced adenine editing rate, yielding a TC-specific base editing tool for TC-to-TT or TC-to-TG conversions that broadens the utility of base editors.
腺嘌呤碱基编辑器（ABEs）可在感兴趣的基因组位点催化特定的A到G的转换。然而，ABEs也会在靶位点诱导胞嘧啶脱氨。为了降低胞嘧啶编辑活性，我们对一种常用的腺苷脱氨酶TadA7.10进行了改造，发现TadA7.10中具有D108Q突变的ABE7.10的胞嘧啶脱氨活性降低了十倍。D108Q突变也降低了最近开发的两种高活性ABE版本ABE8e和ABE8s中的胞嘧啶脱氨活性，并且与降低脱靶RNA编辑的V106W突变兼容。含有P48R突变的ABE7.10表现出增加的胞嘧啶脱氨活性和显著降低的腺嘌呤编辑率，产生了一种用于TC到TT或TC到TG转换的TC特异性碱基编辑工具，拓宽了碱基编辑器的应用范围。
