FLInt: single shot safe harbor transgene integration via Fluorescent Landmark Interference.

ABSTRACT: The stable incorporation of transgenes and recombinant DNA material into the host genome is a bottleneck in many bioengineering applications. Due to the low efficiency, identifying the transgenic animals is often a needle in the haystack. Thus, optimal conditions require efficient screening procedures, but also known and safe landing sites that do not interfere with host expression, low input material and strong expression from the new locus. Here, we leverage an existing library of ≈300 different loci coding for fluorescent markers that are distributed over all 6 chromosomes in Caenorhabditis elegans as safe harbors for versatile transgene integration sites using CRISPR/Cas9. We demonstrated that a single crRNA was sufficient for cleavage of the target region and integration of the transg

SUBMITTER: Malaiwong N 

PROVIDER: S-EPMC10151404 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature