Our lab performs CRISPR injections into embryonic zebrafish using the Alt-R S.p. Cas9 Nuclease (WT Cas9) system. For affinity tagging of genes we generally co-inject small, ~200bp HDR donor oligos (ssDNA) that are not chemically modified with the gene-specific RNP complex (Cas9 enzyme & sgRNA). I would like to find a way to increase the frequency of donor integration in our injections and found the Alt-R HDR Enhancer V2 from IDT. A lab alumnus here has used this compound before during injections, but the actual protocol he used is not clear; safety precautions, toxicity tests, etc. were either not performed or never listed. The protocols for this product on IDT consider only its uses in cultured cells (transfection via nucleofection), and I can't find any evidence of its use in vivo. Has anyone used this compound for Cas9 CRISPR injections? Does it work to increase integration? What should I consider when using this product in an in vivo model system?
Thanks!
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