Published online 24 February 2004
Nucleic Acids Research, 2004, Vol. 32, No. 4 e40
© 2004 Oxford University Press
Homologous gene targeting in Caenorhabditis elegans by biolistic transformation
Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands and 1 Departments of Anatomy and of Biochemistry and Biophysics, Programs in Developmental Biology, Genetics and Neuroscience, Howard Hughes Medical Institute, Box 0452, University of California, San Francisco, CA 94143, USA
*To whom correspondence should be addressed. Tel: +31 30 212 1828; Fax: +31 30 251 6554; Email: plasterk{at}niob.knaw.nl
Targeted homologous recombination is a powerful approach for genome manipulation that is widely used for gene alteration and knockouts in mouse and yeast. In Caenorhabditis elegans, several methods of target-selected mutagenesis have been implemented but none of them provides the opportunity of introducing exact predefined changes into the genome. Although anecdotal cases of homologous gene targeting in C.elegans have been reported, no practical technique of gene targeting has been developed so far. In this work we demonstrate that transformation of C.elegans by microparticle bombardment (biolistic transformation) can result in homologous recombination between introduced DNA and the chromosomal locus. We describe a scaled up version of biolistic transformation that can be used as a method for homologous gene targeting in the worm.
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