Single and dual drug selection for transgenes following bombardment of Caenorhabditis species.
The use of drugs and drug resistance genes is a powerful method to select for the presence of a transgene. Unlike methods that require the complementation of a genetic mutation, this system can be used on any genetic background. Drug selection does not require extensive manipulation or costly equipment, yet it is very rapid and can achieve extremely high efficiency, selecting a small number of transgenic worms from among millions of non-transgenic worms. Introducing integrated transgenes into Caenorhabditis elegans by microparticle bombardment represents just such a challenge. Here we describe in detail the protocol we have developed for dual-drug selection in liquid with puromycin and G418 which works well in a variety of Caenorhabditis species. We also show that single drug selection with only puromycin or only G418 is effective in C. elegans. The growing number of drug selection markers that have been adapted to C. elegans are an important addition to the genetic toolkit at our disposal.
EMBL-CRG Systems Biology Unit, Centre for Genomic Regulation (CRG), Dr. Aiguader 88, 08003 Barcelona, Spain; Universitat Pompeu Fabra (UPF), Dr. Aiguader 88, 08003 Barcelona, Spain. Electronic address: semple.j@gmail.com.
Jennifer I
JI
Semple
EMBL-CRG Systems Biology Unit, Centre for Genomic Regulation (CRG), Dr. Aiguader 88, 08003 Barcelona, Spain; Universitat Pompeu Fabra (UPF), Dr. Aiguader 88, 08003 Barcelona, Spain; Institucio ́Catalana de Recerca i Estudis Avançats (ICREA), Passeig Lluis Companys 23, 08010 Barcelona, Spain.
Ben
B
Lehner
D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology / Q000627:therapeutic use*; D017173:Caenorhabditis elegans / Q000187:drug effects*; D004352:Drug Resistance, Microbial / Q000235:genetics*; D018014:Gene Transfer Techniques; D009154:Mutation; D011691:Puromycin / Q000008:administration & dosage; D019076:Transgenes
D000900:Anti-Bacterial Agents; D011691:Puromycin
Antibiotic; C. brenneri; C. briggsae; C. elegans; C. remanei; Drug selection; G418; Microparticle bombardment; Puromycin; Transgenesis