Transgenesis in model organisms is an essential tool for determining the function of protein-coding genes and non-coding regulatory regions. In Caenorhabditis elegans, injected DNA can be propagated as multicopy extra-chromosomal arrays, but transgenes in arrays are frequently mosaic, over-expressed in some tissues, and silenced in the germline. Here, we describe methods to insert single-copy transgenes into specific genomic locations (MosSCI) or random locations (miniMos) using Mos1 transposons. Single-copy insertions allow expression at endogenous levels, expression in the germline, and identification of active and repressed regions of the genome.
|Original language||English (US)|
|Title of host publication||Methods in Molecular Biology|
|Number of pages||18|
|State||Published - Mar 24 2022|
Bibliographical noteKAUST Repository Item: Exported on 2022-05-26
Acknowledgements: Research in the laboratory of CFJ is supported by KAUST intramural funding. Strains for transgene insertion are provided by the CGC, which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440), and by the NemaGENETAG project. We thank the Bargmann lab (Rockefeller University) for plasmids used for histamine-based selection.