Generation of stable transgenic C. elegans using microinjection.
Transgenic Caenorhabditis elegans can be readily created via microinjection of a DNA plasmid solution into the gonad. The plasmid DNA rearranges to form extrachromosomal concatamers that are stably inherited, though not with the same efficiency as actual chromosome. A gene of interest is co-injected with an obvious phenotypic marker, such as rol-6 or GFP, to allow selection of transgenic animals under a dissecting microscope. The exogenous gene may be expressed from its native promoter for cellular localization studies. Alternatively, the transgene can be driven by a different tissue-specific promoter to assess the role of the gene product in that particular cell or tissue. This technique efficiently drives gene expression in all tissues of C. elegans except for the germline or early embryo. Creation of transgenic animals is widely utilized for a range of experimental paradigms. This video demonstrates the microinjection procedure to generate transgenic worms. Furthermore, selection and maintenance of stable transgenic C. elegans lines is described.
Berkowitz
Laura A
LA
Department of Biological Sciences, University of Alabama, USA. laberkowitz@bama.ua.edu
Knight
Adam L
AL
Caldwell
Guy A
GA
Caldwell
Kim A
KA
D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000235:genetics*; D004247:DNA / Q000235:genetics; D008845:Microinjections / Q000379:methods*; D010957:Plasmids / Q000235:genetics; D019076:Transgenes
D004247:DNA