小柯机器人

近日，荷兰乌得勒支大学医学中心Hans Clevers、Jeffrey M. Beekman等研究人员合作发现，基于CRISPR的腺嘌呤编辑器可纠正囊性纤维化类器官体的无义突变。2020年2月20日，《细胞—干细胞》杂志在线发表了这一成果。

研究人员报道了囊性纤维化（CF）肠类器官体生物库（代表了664名病人），理论上腺嘌呤碱基编辑器（ABE）可以修复其中的20％。研究人员将SpCas9-ABE（PAM识别序列：NGG）和xCas9-ABE（PAM识别序列：NGN）应用于四个选定的CF类器官体样本。在这四例病例中均获得了遗传和功能修复，而两名患者校正品系的全基因组测序（WGS）未检测到脱靶突变。这些观察结果证明了体现患者遗传性疾病的大型患者类器官生物库的价值，并表明ABE可以安全地应用于人体细胞。

据介绍，ABE可实现从A-T到G-C碱基对的酶促转化。ABE具有临床应用前景，因为它不依赖于双链断裂，这与传统的CRISPR/Cas9介导的基因组工程相反。

附：英文原文

Title: CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank

Author: Maarten H. Geurts, Eyleen de Poel, Gimano D. Amatngalim, Rurika Oka, Fleur M. Meijers, Evelien Kruisselbrink, Peter van Mourik, Gitte Berkers, Karin M. de Winter-de Groot, Sabine Michel, Danya Muilwijk, Bente L. Aalbers, Jasper Mullenders, Sylvia F. Boj, Sylvia W.F. Suen, Jesse E. Brunsveld, Hettie M. Janssens, Marcus A. Mall, Simon Y. Graeber, Ruben van Boxtel, Cornelis K. van der Ent, Jeffrey M. Beekman, Hans Clevers

Issue&Volume: 2020-02-20

Abstract: Adenine base editing (ABE) enables enzymatic conversion from A-T into G-C base pairs.ABE holds promise for clinical application, as it does not depend on the introductionof double-strand breaks, contrary to conventional CRISPR/Cas9-mediated genome engineering.Here, we describe a cystic fibrosis (CF) intestinal organoid biobank, representing664 patients, of which ~20% can theoretically be repaired by ABE. We apply SpCas9-ABE(PAM recognition sequence: NGG) and xCas9-ABE (PAM recognition sequence: NGN) on fourselected CF organoid samples. Genetic and functional repair was obtained in all fourcases, while whole-genome sequencing (WGS) of corrected lines of two patients didnot detect off-target mutations. These observations exemplify the value of large,patient-derived organoid biobanks representing hereditary disease and indicate thatABE may be safely applied in human cells.

DOI: 10.1016/j.stem.2020.01.019

Source: https://www.cell.com/cell-stem-cell/fulltext/S1934-5909(20)30019-9

Cell Stem Cell：《细胞—干细胞》，创刊于2007年。隶属于细胞出版社，最新IF：25.269
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