Fly-TILL: Reverse genetics using a living point mutation resource

 Abstract

Mutagenesis with ethylmethanesulfonate (EMS) has been the standard for traditional genetic screens, and in recent years has been applied to reverse genetics. However, reverse-genetic strategies require maintaining a viable germline library so that mutations that are discovered can subsequently be recovered. In applying our TILLING (Targeting Induced Local Lesions IN Genomes) method to establish a Drosophila reverse-genetic service (Fly-TILL), we chose to screen the Zuker lines, a large collection of EMS-mutagenized second- and third-chromosome balanced lines that had been established for forward-genetic screening. For the past four years, our Fly-TILL service has screened this collection to provide ~150 allelic series of point mutations for the fly community. Our analysis of >2000 point mutations and indels has provided a glimpse into the population dynamics of this valuable genetic resource. We found evidence for selection and differential recovery of mutations, depending on distance from balancer breakpoints. Although this process led to variable mutational densities, we have nevertheless been able to deliver valuable mutations in genes selected by Fly-TILL users. We anticipate that our findings will help guide the future implementation of point-mutation resources for the Drosophila community.

 Related Article:

JL Cooper, EA Greene, BJ Till, CA Codomo, BT Wakimoto, S Henikoff. Retention of induced mutations in a Drosophila reverse-genetic resource. Genetics 2008; 180: 661- 7.
PMID: 19015548 DOI: 10.1534/genetics.108

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Fly-TILL: Reverse genetics using a living point mutation resource