Efficient correction of Duchenne muscular dystrophy mutations by SpCas9 and dual gRNAs

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Efficient correction of Duchenne muscular dystrophy mutations by SpCas9 and dual gRNAs. / Xiang, Xi; Zhao, Xiaoying; Pan, Xiaoguang; Dong, Zhanying; Yu, Jiaying; Li, Siyuan; Liang, Xue; Han, Peng; Qu, Kunli; Jensen, Jonas Brorson; Farup, Jean; Wang, Fei; Petersen, Trine Skov; Bolund, Lars; Teng, Huajing; Lin, Lin; Luo, Yonglun.

In: Molecular Therapy - Nucleic Acids, Vol. 24, 2021, p. 403-415.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Xiang, X, Zhao, X, Pan, X, Dong, Z, Yu, J, Li, S, Liang, X, Han, P, Qu, K, Jensen, JB, Farup, J, Wang, F, Petersen, TS, Bolund, L, Teng, H, Lin, L & Luo, Y 2021, 'Efficient correction of Duchenne muscular dystrophy mutations by SpCas9 and dual gRNAs', Molecular Therapy - Nucleic Acids, vol. 24, pp. 403-415. https://doi.org/10.1016/j.omtn.2021.03.005

APA

Xiang, X., Zhao, X., Pan, X., Dong, Z., Yu, J., Li, S., Liang, X., Han, P., Qu, K., Jensen, J. B., Farup, J., Wang, F., Petersen, T. S., Bolund, L., Teng, H., Lin, L., & Luo, Y. (2021). Efficient correction of Duchenne muscular dystrophy mutations by SpCas9 and dual gRNAs. Molecular Therapy - Nucleic Acids, 24, 403-415. https://doi.org/10.1016/j.omtn.2021.03.005

Vancouver

Xiang X, Zhao X, Pan X, Dong Z, Yu J, Li S et al. Efficient correction of Duchenne muscular dystrophy mutations by SpCas9 and dual gRNAs. Molecular Therapy - Nucleic Acids. 2021;24:403-415. https://doi.org/10.1016/j.omtn.2021.03.005

Author

Xiang, Xi ; Zhao, Xiaoying ; Pan, Xiaoguang ; Dong, Zhanying ; Yu, Jiaying ; Li, Siyuan ; Liang, Xue ; Han, Peng ; Qu, Kunli ; Jensen, Jonas Brorson ; Farup, Jean ; Wang, Fei ; Petersen, Trine Skov ; Bolund, Lars ; Teng, Huajing ; Lin, Lin ; Luo, Yonglun. / Efficient correction of Duchenne muscular dystrophy mutations by SpCas9 and dual gRNAs. In: Molecular Therapy - Nucleic Acids. 2021 ; Vol. 24. pp. 403-415.

Bibtex

@article{8b1ee1cf1ffa44e19807e72fb7b9bf41,
title = "Efficient correction of Duchenne muscular dystrophy mutations by SpCas9 and dual gRNAs",
abstract = "CRISPR gene therapy is one promising approach for treatment of Duchenne muscular dystrophy (DMD), which is caused by a large spectrum of mutations in the dystrophin gene. To broaden CRISPR gene editing strategies for DMD treatment, we report the efficient restoration of dystrophin expression in induced myotubes by SpCas9 and dual guide RNAs (gRNAs). We first sequenced 32 deletion junctions generated by this editing method and revealed that non-homologous blunt-end joining represents the major indel type. Based on this predictive repair outcome, efficient in-frame deletion of a part of DMD exon 51 was achieved in HEK293T cells with plasmids expressing SpCas9 and dual gRNAs. More importantly, we further corrected a frameshift mutation in human DMD (exon45del) fibroblasts with SpCas9-dual gRNA ribonucleoproteins. The edited DMD fibroblasts were transdifferentiated into myotubes by lentiviral-mediated overexpression of a human MYOD transcription factor. Restoration of DMD expression at both the mRNA and protein levels was confirmed in the induced myotubes. With further development, the combination of SpCas9-dual gRNA-corrected DMD patient fibroblasts and transdifferentiation may provide a valuable therapeutic strategy for DMD.",
author = "Xi Xiang and Xiaoying Zhao and Xiaoguang Pan and Zhanying Dong and Jiaying Yu and Siyuan Li and Xue Liang and Peng Han and Kunli Qu and Jensen, {Jonas Brorson} and Jean Farup and Fei Wang and Petersen, {Trine Skov} and Lars Bolund and Huajing Teng and Lin Lin and Yonglun Luo",
year = "2021",
doi = "10.1016/j.omtn.2021.03.005",
language = "English",
volume = "24",
pages = "403--415",
journal = "Molecular Therapy - Nucleic Acids",
issn = "2162-2531",
publisher = "nature publishing group",

}

RIS

TY - JOUR

T1 - Efficient correction of Duchenne muscular dystrophy mutations by SpCas9 and dual gRNAs

AU - Xiang, Xi

AU - Zhao, Xiaoying

AU - Pan, Xiaoguang

AU - Dong, Zhanying

AU - Yu, Jiaying

AU - Li, Siyuan

AU - Liang, Xue

AU - Han, Peng

AU - Qu, Kunli

AU - Jensen, Jonas Brorson

AU - Farup, Jean

AU - Wang, Fei

AU - Petersen, Trine Skov

AU - Bolund, Lars

AU - Teng, Huajing

AU - Lin, Lin

AU - Luo, Yonglun

PY - 2021

Y1 - 2021

N2 - CRISPR gene therapy is one promising approach for treatment of Duchenne muscular dystrophy (DMD), which is caused by a large spectrum of mutations in the dystrophin gene. To broaden CRISPR gene editing strategies for DMD treatment, we report the efficient restoration of dystrophin expression in induced myotubes by SpCas9 and dual guide RNAs (gRNAs). We first sequenced 32 deletion junctions generated by this editing method and revealed that non-homologous blunt-end joining represents the major indel type. Based on this predictive repair outcome, efficient in-frame deletion of a part of DMD exon 51 was achieved in HEK293T cells with plasmids expressing SpCas9 and dual gRNAs. More importantly, we further corrected a frameshift mutation in human DMD (exon45del) fibroblasts with SpCas9-dual gRNA ribonucleoproteins. The edited DMD fibroblasts were transdifferentiated into myotubes by lentiviral-mediated overexpression of a human MYOD transcription factor. Restoration of DMD expression at both the mRNA and protein levels was confirmed in the induced myotubes. With further development, the combination of SpCas9-dual gRNA-corrected DMD patient fibroblasts and transdifferentiation may provide a valuable therapeutic strategy for DMD.

AB - CRISPR gene therapy is one promising approach for treatment of Duchenne muscular dystrophy (DMD), which is caused by a large spectrum of mutations in the dystrophin gene. To broaden CRISPR gene editing strategies for DMD treatment, we report the efficient restoration of dystrophin expression in induced myotubes by SpCas9 and dual guide RNAs (gRNAs). We first sequenced 32 deletion junctions generated by this editing method and revealed that non-homologous blunt-end joining represents the major indel type. Based on this predictive repair outcome, efficient in-frame deletion of a part of DMD exon 51 was achieved in HEK293T cells with plasmids expressing SpCas9 and dual gRNAs. More importantly, we further corrected a frameshift mutation in human DMD (exon45del) fibroblasts with SpCas9-dual gRNA ribonucleoproteins. The edited DMD fibroblasts were transdifferentiated into myotubes by lentiviral-mediated overexpression of a human MYOD transcription factor. Restoration of DMD expression at both the mRNA and protein levels was confirmed in the induced myotubes. With further development, the combination of SpCas9-dual gRNA-corrected DMD patient fibroblasts and transdifferentiation may provide a valuable therapeutic strategy for DMD.

U2 - 10.1016/j.omtn.2021.03.005

DO - 10.1016/j.omtn.2021.03.005

M3 - Journal article

C2 - 33868784

VL - 24

SP - 403

EP - 415

JO - Molecular Therapy - Nucleic Acids

JF - Molecular Therapy - Nucleic Acids

SN - 2162-2531

ER -

ID: 272128383