My lab is embarking on using CRISPR coupled with HDR for transgenic insertion. We have been using CRISPR for gene knock-in for a couple of years now with good success rate (60-90%). However, we know that targeted gene insertion using CRISPR has a much lower success rate.
We know one of major obstacles is preventing NHEJ. We are also planning on using circular donor plasmids, but may also test out single-stranded donor oligos.
So, I was wondering if anyone had any suggestions on how to increase HDR or how to increase the rate of insertion in general?
Dear Max R. Bangs,
Do you happen to know a recent publication about using 'Double Cut' HDR donor (plasmid) to increase CRISPR/Cas9-meidated transgne insertion (through HDR).
Please see an update I wrote at (copy and paste to view):
https://www.researchgate.net/project/collaboration-Working-on-a-scientific-manuscript-about-revolutionary-genome-editing-technology-on-generating-designer-crops-and-beyond?_sg=fmck1qhZ839T23QDYoY_a1B_ogodF5XsaSauiofY5gjlcwnzOXgnNKSe9QHC8vctE3c13CvwCSqPLe9FJMGaM-lzD0ufxZ6h-q7h
- Badges
- Science topic
- Similar topics
- Mathematical Sciences
- Graphs