Using novel TALEN technology to engineer precise mutations in the genome of Drosophila melanogaster
TALENs (transcription activator-like effector nucleases) are DNA-binding proteins that, upon dimerization of the nuclease, cleave their DNA targets. Their modular assembly makes them able to bind virtually any DNA sequence, allowing for their use as a tool in genome modification. TALEN technology emerged recently in molecular biology, as they were characterized just two years ago, and their use has been largely unexplored in Drosophila melanogaster. Published data has demonstrated the use of TALENs to excise small, targeted pieces of the fly genome. My research project aims to express TALENs in the fly genome (which has not been done before) and to use them to target a foreign loxP target sequence in the genome for excision. The loxP site is a synthetic byproduct of homologous recombination, a technique widely used in our lab for precisely engineering in vivo genetic mutations; for example the introduction of a disease-causing mutation into a target gene. LoxP sequences flank a red eye gene in Drosophila and removal or disruption of the loxP site can easily be scored by the loss of red pigment in the eyes. Successful TALEN expression in flies will not only further the understanding of TALEN use in Drosophila but will also report on whether or not TALEN expression from the genome, rather than injecting mRNA coding for the TALEN protein, is a feasible strategy for Drosophila genetic engineering.
A file with the transcript of the narration can be found here