Generating a grin1a mutation in Zebrafish

• In Event: Poster Presentation and Judging for: Anthropology; Botany; Conservation Ecology; Environmental Science; Freshwater Science; Geosciences; Mathematics & Computer Science; Neuroscience; Physics & Eng; Science Education; Terrestrial Ecology & Management.
  In Poster Session: Neuroscience Poster Session

Abstract

The gene grin1a encodes the NR1.1 subunit of the NMDA ionotropic glutamate receptor. This receptor plays fundamental role in the molecular processes of learning and memory in many animals. Furthermore, mutations and polymorphisms in this gene have been associated with schizophrenia. Animal models greatly facilitate the analysis of gene functionality; unfortunately mice with null mutations die prenatally. The zebrafish provides a unique opportunity, as a partial genome duplication led to two GRIN1 homologues, (grin1a and grin1b). The goal of this work is to utilize the CRISPR/Cas9 system to generate a grin1a null mutant in zebrafish. In addition, to facilitate rapid genotyping, we will replace the third exon with a blue fluorescent protein. We designed potential guide RNAs, synthesized them using T4 polymerase primer extension, and transcribed the guides and Cas9 using in vitro transcription. Initial injections of the CRISPR/Cas9 RNAs have been initiated, and we are currently screening for mutations by PCR. Once mutations are identified, phenotyping will begin at the morphologic and behavioral level.

Authors

• Sahir Burkie, The university of Texas at Tyler

• Brent R Bill, University of Texas at Tyler