[PDF][PDF] Correction of a genetic disease in mouse via use of CRISPR-Cas9

Y Wu, D Liang, Y Wang, M Bai, W Tang, S Bao, Z Yan… - Cell stem cell, 2013 - cell.com
Y Wu, D Liang, Y Wang, M Bai, W Tang, S Bao, Z Yan, D Li, J Li
Cell stem cell, 2013cell.com
The CRISPR-Cas9 system has been employed to generate mutant alleles in a range of
different organisms. However, so far there have not been reports of use of this system for
efficient correction of a genetic disease. Here we show that mice with a dominant mutation in
Crygc gene that causes cataracts could be rescued by coinjection into zygotes of Cas9
mRNA and a single-guide RNA (sgRNA) targeting the mutant allele. Correction occurred via
homology-directed repair (HDR) based on an exogenously supplied oligonucleotide or the …
Summary
The CRISPR-Cas9 system has been employed to generate mutant alleles in a range of different organisms. However, so far there have not been reports of use of this system for efficient correction of a genetic disease. Here we show that mice with a dominant mutation in Crygc gene that causes cataracts could be rescued by coinjection into zygotes of Cas9 mRNA and a single-guide RNA (sgRNA) targeting the mutant allele. Correction occurred via homology-directed repair (HDR) based on an exogenously supplied oligonucleotide or the endogenous WT allele, with only rare evidence of off-target modifications. The resulting mice were fertile and able to transmit the corrected allele to their progeny. Thus, our study provides proof of principle for use of the CRISPR-Cas9 system to correct genetic disease.
cell.com