Knock-ins are hard, for a long time people even said they were impossible, but we are nearing an 80% success rate for CRISPR knock-ins in zebrafish so we want to share with you what we believe to be the 11 steps necessary to have a successful knock-in project.
A client wanted a knockout (KO) of an embryonic lethal gene. We could not make this line using our standard methods. Instead, we inserted two loxP sites. One in the first intron of the gene and the second in the 3’utr. After we confirmed this line by PCR and sequencing, we injected this line with …
Insertion of two loxP sites to produce a knockout (KO) of an embryonic lethal gene Read More »
