title: 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.
authors.affiliation: Graduate School of the Biomedical Sciences, Rowan University.
authors.forename: Qing
authors.initials: Q
authors.lastname: Wei
authors.forename: Yongquan
authors.initials: Y
authors.lastname: Shen
authors.forename: Xiangmei
authors.initials: X
authors.lastname: Chen
authors.forename: Yelena
authors.initials: Y
authors.lastname: Shifman
authors.forename: Ronald E
authors.initials: RE
authors.lastname: Ellis
mesh_terms: D000818:Animals; D001483:Base Sequence; D005075:Biological Evolution; D002107:Caenorhabditis / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D019295:Computational Biology; D004720:Endonucleases / Q000378:metabolism*; D055786:Gene Knockout Techniques; D009154:Mutation; D013045:Species Specificity; D015534:Trans-Activators / Q000378:metabolism*
chemical_list: D015534:Trans-Activators; D004720:Endonucleases
keywords: C. briggsae; TALENs; evolution; model organism; nematode