PhiC31 integrase for recombination-mediated single-copy insertion and genome manipulation in Caenorhabditis elegans.

ABSTRACT: Caenorhabditis elegans benefits from a large set of tools for genome manipulation. Yet, the precise single-copy insertion of very large DNA constructs (>10 kb) and the generation of inversions are still challenging. Here, we adapted the phiC31 integrase system for C. elegans. We generated an integrated phiC31 integrase expressing strain flanked by attP sites that serves as a landing pad for integration of transgenes by recombination-mediated cassette exchange (RCME). This strain is unc-119(-) so RMCE integrants can be produced simply by injection of a plasmid carrying attB sites flanking unc-119(+) and the gene(s) of interest. Additionally, phiC31 integrase is removed concomitantly with integration, eliminating the need to outcross away the integrase. Integrations were obtained for insert

SUBMITTER: Yang FJ 

PROVIDER: S-EPMC9208643 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature