- Target selection is free from PAM sequence constraints.
- Ex-vivo therapy: realized by transfection of RNA or protein.
- In-vivo therapy: Components are small enough for AAV.
Although gene disruption therapy by CRISPR is actively being developed, major challenges of CRISPR to use as a tool for gene correction are to : i) ensure homology-directed repair (HDR), precise genetic change without insertion or deletion (indels) at the target site and ii) avoid off-target mutations, without scarifying gene correction efficiency.
Recently, base editing technologies were invented, in which deaminase linked with Cas9 nickase alters C to T, or A to G, but its application is limited and off-target error still exist.
Previously, we invented Single Nicking of the target Gene and Donor(SNGD) plasmid -induced surprisingly precise and efficient gene correction method (doi:10.1101/gr.226027.117). In this technology, we have taken one step further, utilized the other allele as a donner. Given that mutation points are different in two alleles of most recessive genetic disorder patients, this strategy is applicable for many of them.
Facts and Data
- Thymidine kinase (TK) Point mutation of TK6 derived TK deficiency lymphoblast cells were corrected up to 5% by electroporation of RNA. Very few indels or off target editing were observed.
- fibroblast cells from hereditary genome instability syndrome were gene corrected and showed normal protein expression.
Patents and Publications
- Journal in preparation
Prof. Shinichiro Nakada, Department of Bioregulation and Cellular Response, Graduate School of Medicine, Osaka Univ., Japan
We are seeking licensees who utilize this technology for gene therapy. Collaborative researches targeting specific diseases are welcome. Details can be disclosed under CDA.
Product No. KJ-02315