Abstract
Mutagenesis drives natural selection. In the lab, mutations allow gene function to be deciphered. C. elegans is highly amendable to functional genetics because of its short generation time, ease of use, and wealth of available gene-alteration techniques. Here we provide an overview of historical and contemporary methods for mutagenesis in C. elegans, and discuss principles and strategies for forward (genome-wide mutagenesis) and reverse (target-selected and gene-specific mutagenesis) genetic studies in this animal.
Highlights
Mutations are the engine of natural selection and arise spontaneously through a myriad of mechanisms (Nei, 2007)
Non-directed genome-wide mutagenesis is commonly used for forward genetic screens, where mutants defective in a biological process of interest are sought
Mutations are generated at rates of 2.8 × 10-2 to 3.9 × 10-3 per haploid genome. 32P is incorporated uniformly into genomic DNA, and mutation is caused by 32P-transmutation in the genome, rather than by β-particle emission (Babu and Brenner, 1981). 32P is mixed with unlabeled phosphate and added directly to phosphate-free NGM plates at 10 Ci/mol! The plates are seeded with NA22, a radioactive-resistant strain of bacteria (Anderson, 1995)
Summary
Mutations are the engine of natural selection and arise spontaneously through a myriad of mechanisms (Nei, 2007). In C. elegans, spontaneous point mutations, gene duplications, and deletions arise at rates of 2 × 10-8, 3.4 × 10-7, and 2.2 × 10-7 per gene per generation, respectively (Denver et al, 2004; Lipinski et al, 2011). These alterations to the genome result from replication and repair errors, endogenous oxidative damage, and exogenous environmental insults (Denver et al, 2009). For spontaneous mutations to be an effective tool, roughly 1/(mutation rate) organisms must be screened to identify lesions in a particular gene. Make targeted changes or deletions in gene of interest without mutagenizing entire genome
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
