Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
Key Lab of Computational Biology, CAS-MPG Partner Institute for Computational Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
Genome Biol. 2017 Nov 24;18(1):224. doi: 10.1186/s13059-017-1354-4.
The CRISPR/Cas9 system has become an efficient gene editing method for generating cells carrying precise gene mutations, including the rearrangement and deletion of chromosomal segments. However, whether an entire chromosome could be eliminated by this technology is still unknown.
Here we demonstrate the use of the CRISPR/Cas9 system to eliminate targeted chromosomes. Using either multiple cleavages induced by a single-guide RNA (sgRNA) that targets multiple chromosome-specific sites or a cocktail of multiple sgRNAs, each targeting one specific site, we found that a sex chromosome could be selectively eliminated in cultured cells, embryos, and tissues in vivo. Furthermore, this approach was able to produce a targeted autosome loss in aneuploid mouse embryonic stem cells with an extra human chromosome and human induced pluripotent stem cells with trisomy 21, as well as cancer cells.
CRISPR/Cas9-mediated targeted chromosome elimination offers a new approach to develop animal models with chromosome deletions, and a potential therapeutic strategy for human aneuploidy diseases involving additional chromosomes.
CRISPR/Cas9 系统已成为一种高效的基因编辑方法，可用于产生携带精确基因突变的细胞，包括染色体片段的重排和缺失。然而，该技术是否能完全消除整条染色体尚不清楚。
本研究中，我们利用 CRISPR/Cas9 系统来消除靶向染色体。通过使用靶向多个染色体特异性位点的单个向导 RNA（sgRNA）诱导的多次切割，或使用靶向一个特定位点的多个 sgRNA 的混合物，我们发现可以在培养细胞、胚胎和体内组织中选择性地消除性染色体。此外，该方法还能够在具有额外人类染色体的非整倍体小鼠胚胎干细胞和 21 三体人诱导多能干细胞以及癌细胞中产生靶向常染色体缺失。
CRISPR/Cas9 介导的靶向染色体消除为开发具有染色体缺失的动物模型提供了一种新方法，也为涉及额外染色体的人类非整倍体疾病提供了一种潜在的治疗策略。
