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 Evoljournal_title
Molecular biology and evolutionauthors
Wei Q,Shen Y,Chen X,Shifman Y,Ellis REdoi
10.1093/molbev/mst213subject
Has Abstractpub_date
2014-02-01 00:00:00pages
468-73issue
2eissn
0737-4038issn
1537-1719pii
mst213journal_volume
31pub_type
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