Template-independent genome editing and repairing correct frameshift disease in vivo

Liu, L.; Li, K.; Zou, L.; Hou, H.; Hu, Q.; Liu, S.; Wang, S.; Wang, Y.; Li, J.; Song, C.; Chen, J.; Li, C.; Du, H.; Li, J.-L.; Chen, F.; Xu, Z.; Sun, W.; Sun, Q.; Xiong, W.

doi:10.1101/2020.11.13.381160

bioRxiv · 10.1101/2020.11.13.381160

Template-independent genome editing and repairing correct frameshift disease in vivo

Liu, L.·Li, K.·Zou, L.·Hou, H.·Hu, Q.·Liu, S.·Wang, S.·Wang, Y.·Li, J.·Song, C.·Chen, J.·Li, C.·Du, H.·Li, J.-L.·Chen, F.·Xu, Z.·Sun, W.·Sun, Q.·Xiong, W.

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Abstract

Frameshift mutation caused by small insertions/deletions (indels) often generate truncated and non-functional proteins, which underlies 22% inherited Mendelian disorders in humans. However, there is no efficient in vivo gene therapy strategies available to date, especially in postmitotic systems. Here, we leveraged the non-homologous end joining (NHEJ) mediated non-random editing profiles to compensate the frameshift mutation in a USH1F mouse model - av3j. After treatment by the selected gRNA, about 50% editing products showed reading-frame restoration, and more than 70% targeted hair cells recovered mechanotransduction. In vivo treatment ameliorated the hearing and balance symptoms in homozygous mutant mice. Furthermore, a scale-up analysis of 114 gRNAs targeting 40 frameshift deafness mutations reveals that 65% loci have at least one gRNA with predicted therapeutic potential. Together, our study demonstrates that the NHEJ-mediated frame restoration is a simple and highly efficient therapeutic strategy for small-indel induced frameshift mutations.

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Liu, L., Li, K., Zou, L., Hou, H., Hu, Q., Liu, S., Wang, S., Wang, Y., Li, J., Song, C., Chen, J., Li, C., Du, H., Li, J.-L., Chen, F., Xu, Z., Sun, W., Sun, Q., Xiong, W.. 2020-11-15. Template-independent genome editing and repairing correct frameshift disease in vivo. https://doi.org/10.1101/2020.11.13.381160

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