Core Benefit
- Mutation of somatic cells can be corrected with 50% efficiency, not causing NHEJ in another allele, without performing selection.
- Even high rate of bi-allelic gene corrections can be achieved.
Background and Technology
Major challenges of CRISPR to use as a tool for gene therapy are: i) to ensure homology-directed repair (HDR), precise genetic change without inducing additional genetic lesions at the target site and ii) avoiding off-target cleavages. Although single-strand nick repair retain genome integrity, the rate of genetic repair is low, making it unsuitable for gene therapy.
Our proprietary method, single nicking of the target gene and donor plasmid (SNGD)-induced gene correction, details can be disclosed under CDA, is surprisingly more efficient and precise than Cas9-double strand break induced gene correction. Furthermore, the efficiency of nucleotide substitution can markedly increase by consecutive SNGD procedure, with very few errors. The procedure also results in efficient gene correction in both the copies of gene, and even bi-allelic gene corrections can be achieved using SNGD.
Facts and Data
- Point mutation in 293T was corrected by electroporation of Cas9D10A, sgRNA, and donner DNA. Single treatment corrected 18.1% of at least one allele of the cells, in which 14.3% of both copies, only 3.8% indels of the other copy.
- Three consecutive treatment achieved 34% gene correction, in which 33% bi-allele correction, 7 % indels on the other allele. Fourth round increased correction rate to 40%.
- Effect was also confirmed with Hela cells, and endogenous loci.
Patents and Publications
- JP2018-011525
- Genome Res. 2017. doi:10.1101/gr.226027.117
Researcher
Prof. Shinichiro Nakada, Department of Bioregulation and Cellular Response, Graduate School of Medicine, Osaka Univ., Japan
Expectations
We are seeking licensees who utilize this technology for gene therapy or research purpose. Exclusive license can be negotiable. Draft of paper can be disclosed under CDA.
Product No: TP-00630