Rapid generation of Caenorhabditis elegans single-copy transgenes combining recombination-mediated cassette exchange and drug selection.

ABSTRACT: I outline a streamlined method to insert large, single-copy transgenes into the Caenorhabditis elegans genome using recombination-mediated cassette exchange (RMCE) that relies solely on drug selection yielding a homozygous fluorescent protein (FP) marked transgene in 3 generations (8 days) at high efficiency (>1 insertion per 2 injected P0 animals). Landing sites for this approach are available on four chromosomes in several configurations which yield lines marked in distinct cell types. An array of vectors permit creating transgenes using a variety of selection methods (HygR, NeoR, PuroR, and unc-119) that yield lines expressing different colored FPs (BFP, GFP, mNG, and Scarlet). Although these transgenes retain a plasmid backbone and a selection marker, the inclusion of these sequences t

SUBMITTER: Nonet ML 

PROVIDER: S-EPMC10324946 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature