Template-independent genome editing in the Pcdh15av−3j mouse, a model of human DFNB23 nonsyndromic deafness

Liu，Lian1,2; Zou，Linzhi1,2; Li，Kuan1,3; Hou，Hanqing1,2; Hu，Qun1,2; Liu，Shuang1,2; Li，Jie1,2; Song，Chenmeng1,2; Chen，Jiaofeng1,2; Wang，Shufeng1,2; Wang，Yangzhen1,2; Li，Changri4; Du，Haibo5; Li，Jun Liszt6,7; Chen，Fangyi4; Xu，Zhigang5; Sun，Wenzhi6,8; Sun，Qianwen1,3; Xiong，Wei1,2

2022-07-12

10.1016/j.celrep.2022.111061

Cell Reports   Impact Factor & Quartile

2211-1247

2211-1247

Although frameshift mutations lead to 22% of inherited Mendelian disorders in humans, there is no efficient in vivo gene therapy strategy available to date, particularly in nondividing cells. Here, we show that nonhomologous end-joining (NHEJ)-mediated nonrandom editing profiles compensate the frameshift mutation in the Pcdh15 gene and restore the lost mechanotransduction function in postmitotic hair cells of Pcdh15 mice, an animal model of human nonsyndromic deafness DFNB23. Identified by an ex vivo evaluation system in cultured cochlear explants, the selected guide RNA restores reading frame in approximately 50% of indel products and recovers mechanotransduction in more than 70% of targeted hair cells. In vivo treatment shows that half of the animals gain improvements in auditory responses, and balance function is restored in the majority of injected mutant mice. These results demonstrate that NHEJ-mediated reading-frame restoration is a simple and efficient strategy in postmitotic systems.

CP: Molecular biology CP: Neuroscience CRISPR-Cas9 frame restoration gene therapy hair cell in vivo inner ear mechanotransduction nonhomologous end-joining Pcdh15 tip link

SCI

English

Others

National Natural Science Foundation of China[31522025];National Natural Science Foundation of China[31571080];National Natural Science Foundation of China[31861163003];National Natural Science Foundation of China[81873703];

Cell Biology

Cell Biology

WOS:000890459200004

CELL PRESS

2-s2.0-85133890696

Scopus

1.School of Life Sciences,Tsinghua University,Beijing,100084,China
2.IDG/McGovern Institute for Brain Research at Tsinghua University,Tsinghua University,Beijing,100084,China
3.Tsinghua-Peking Joint Center for Life Sciences,Tsinghua University,Beijing,100084,China
4.Department of Biomedical Engineering,Southern University of Science and Technology,Shenzhen,Guangdong,518055,China
5.School of Life Sciences,Shandong University,Qingdao,Shandong,266237,China
6.Chinese Institute for Brain Research,Beijing,102206,China
7.Academy for Advanced Interdisciplinary Studies (AAIS),Peking University,Beijing,100871,China
8.School of Basic Medical Sciences,Capital Medical University,Beijing,100069,China

Liu，Lian,Zou，Linzhi,Li，Kuan,et al. Template-independent genome editing in the Pcdh15av−3j mouse, a model of human DFNB23 nonsyndromic deafness[J]. Cell Reports,2022,40(2).

Liu，Lian.,Zou，Linzhi.,Li，Kuan.,Hou，Hanqing.,Hu，Qun.,...&Xiong，Wei.(2022).Template-independent genome editing in the Pcdh15av−3j mouse, a model of human DFNB23 nonsyndromic deafness.Cell Reports,40(2).

Liu，Lian,et al."Template-independent genome editing in the Pcdh15av−3j mouse, a model of human DFNB23 nonsyndromic deafness".Cell Reports 40.2(2022).