Homologous gene targeting in Caenorhabditis elegans by biolistic transformation.
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.
Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands.
Eugene
E
Berezikov
Cornelia I
CI
Bargmann
Ronald H A
RH
Plasterk
D000818:Animals; D019470:Biolistics / Q000379:methods*; D015139:Blotting, Southern; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics; D002148:Calmodulin-Binding Proteins / Q000235:genetics; D018390:Gene Targeting / Q000379:methods*; D017238:Genes, Helminth / Q000235:genetics; D009124:Muscle Proteins / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D014170:Transformation, Genetic
D029742:Caenorhabditis elegans Proteins; D002148:Calmodulin-Binding Proteins; D009124:Muscle Proteins; C489233:unc-22 protein, C elegans