Versatile strategy for isolating transcription activator‐like effector nuclease‐mediated knockout mutants in Caenorhabditis elegans. (17th December 2013)

• Record Type: Journal Article
• Authors: Sugi, Takuma; Sakuma, Tetsushi; Ohtani, Yasuko; Yamamoto, Takashi
• Is Part Of: Development growth and differentiation. Volume 56:Number 1(2014)
• Page Start: 78
• Page End: 85
• Publication Date: 2013-12-17
• Subjects: Embryology -- Periodicals; Developmental biology -- Periodicals; Growth -- Periodicals; 574.3
• DOI: 10.1111/dgd.12108
• Languages: English
• ISSNs: 0012-1592

Abstract

<abstract abstract-type="main" id="dgd12108-abs-0001"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Targeted genome editing using transcription activator‐like effector nuclease (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 systems has recently emerged as a potentially powerful method for creating locus‐specific mutations in <italic>Caenorhabditis elegans</italic>. Due to the low mutation frequencies, one of the crucial steps in using these technologies is screening animals that harbor a targeted mutation. In previous studies, identifying targeted mutations in <italic>C. elegans</italic> usually depended on observations of fluorescent markers such as a green fluorescent protein or visible phenotypes such as dumpy and uncoordinated phenotypes. However, this strategy is limited in practice because the phenotypes caused by targeted mutations such as defects in sensory behaviors are often apparently invisible. Here, we describe a versatile strategy for isolating <italic>C. elegans</italic> knockout mutants by TALEN‐mediated genome editing and a heteroduplex mobility assay. We applied TALENs to engineer the locus of the neural gene glr‐1, which is a <italic>C. elegans </italic>AMPA‐type receptor orthologue that is known to have crucial roles in various sensory behaviors. Knockout mutations in the <italic>glr‐1</italic> locus, which caused defective mechanosensory behaviors, were efficiently identified by the heteroduplex mobility assay. Thus, we demonstrated the utility of a TALEN‐based knockout strategy for creating <italic>C. elegans</italic> with mutations that cause invisible phenotypes.</p> </abstract>