Therapeutic Genome Editing for Myotonic Dystrophy Type 1 Using CRISPR/Cas9.
Therapeutic Genome Editing for Myotonic Dystrophy Type 1 Using CRISPR/Cas9.
复制标题
DOI：
10.1016/j.ymthe.2018.09.003
复制
发表时间：
2018-11-07
期刊：
影响因子：
--
通讯作者：
Xia G
中科院分区：
文献类型：
--
作者：
Wang Y;Hao L;Wang H;Santostefano K;Thapa A;Cleary J;Li H;Guo X;Terada N;Ashizawa T;Xia G
强直性肌营养不良1型（DM 1）是由强直性肌营养不良症蛋白激酶基因3′ UTR内的CTG核苷酸重复扩增引起的。在这项研究中，我们探索了使用CRISPR/Cas9的治疗性基因组编辑，通过靶向删除扩展的CTG重复序列，并在CTG重复序列的3′ UTR上游靶向插入多聚腺苷酸化信号，以消除有毒的RNA CUG重复序列。我们发现配对的SpCas 9或SaCas 9引导RNA诱导了扩增的CTG重复序列的缺失。然而，这种方法在突变体和正常等位基因中都引起了频繁的倒位。相比之下，在CTG重复序列上游的3 'UTR中插入多聚腺苷酸化信号消除了有毒的RNA CUG重复序列，从而导致分化的神经干细胞、前脑神经元、心肌细胞和骨骼肌肌纤维的表型逆转。我们得出结论，在3′ UTR中靶向插入多聚腺苷酸化信号是开发DM 1治疗性基因组编辑的可行方法。强直性肌营养不良1型是由DMPK基因3′ UTR中具有扩展的CUG重复序列的毒性RNA引起的。Wang等人开发了一种策略，通过在扩增的重复序列上游插入聚腺苷酸化信号来消除毒性重复序列，用于个性化的基于细胞的治疗和体内治疗性基因组编辑。
Myotonic dystrophy type 1 (DM1) is caused by a CTG nucleotide repeat expansion within the 3′ UTR of the Dystrophia Myotonica protein kinase gene. In this study, we explored therapeutic genome editing using CRISPR/Cas9 via targeted deletion of expanded CTG repeats and targeted insertion of polyadenylation signals in the 3′ UTR upstream of the CTG repeats to eliminate toxic RNA CUG repeats. We found paired SpCas9 or SaCas9 guide RNA induced deletion of expanded CTG repeats. However, this approach incurred frequent inversion in both the mutant and normal alleles. In contrast, the insertion of polyadenylation signals in the 3′ UTR upstream of the CTG repeats eliminated toxic RNA CUG repeats, which led to phenotype reversal in differentiated neural stem cells, forebrain neurons, cardiomyocytes, and skeletal muscle myofibers. We concluded that targeted insertion of polyadenylation signals in the 3′ UTR is a viable approach to develop therapeutic genome editing for DM1. Myotonic dystrophy type 1 is caused by toxic RNAs with expanded CUG repeats in the 3′ UTR of the DMPK gene. Wang et al. developed a strategy to eliminate the toxic repeats by insertion of polyadenylation signals upstream of the expanded repeats for personalized cell-based therapy and in vivo therapeutic genome editing.
登录
查看更多内容
影响因子：
7
作者：
Cho SW;Kim S;Kim Y;Kweon J;Kim HS;Bae S;Kim JS
通讯作者：
Kim JS
影响因子：
16.2
作者：
Ash PE;Bieniek KF;Gendron TF;Caulfield T;Lin WL;Dejesus-Hernandez M;van Blitterswijk MM;Jansen-West K;Paul JW 3rd;Rademakers R;Boylan KB;Dickson DW;Petrucelli L
通讯作者：
Petrucelli L
影响因子：
3.4
作者：
HUARD, J;BOUCHARD, JP;TREMBLAY, JP
通讯作者：
TREMBLAY, JP
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
2.8
作者：
Furling, D;Lemieux, D;Puymirat, J
通讯作者：
Puymirat, J