Rapid creation of forward-genetics tools for C. briggsae using TALENs: lessons for nonmodel organisms.

Abstract:

:Although evolutionary studies of gene function often rely on RNA interference, the ideal approach would use reverse genetics to create null mutations for cross-species comparisons and forward genetics to identify novel genes in each species. We have used transcription activator-like effector nucleases (TALENs) to facilitate both approaches in Caenorhabditis nematodes. First, by combining golden gate cloning and TALEN technology, we can induce frameshifting mutations in any gene. Second, by combining this approach with bioinformatics we can predict and create the resources needed for forward genetic analysis in species like Caenorhabditis briggsae. Although developing genetic model organisms used to require years to isolate marker mutations, balancers, and tools, with TALENs, these reagents can now be produced in months. Furthermore, the analysis of nonsense mutants in related model organisms allows a directed approach for making these markers and tools. When used together, these methods could simplify the adaptation of other organisms for forward and reverse genetics.

journal_name

Mol Biol Evol

authors

Wei Q,Shen Y,Chen X,Shifman Y,Ellis RE

doi

10.1093/molbev/mst213

subject

Has Abstract

pub_date

2014-02-01 00:00:00

pages

468-73

issue

2

eissn

0737-4038

issn

1537-1719

pii

mst213

journal_volume

31

pub_type

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