College of Arts and Science
Ph.D, Quantitative Biology, University of Texas at Arlington
B.Tech, Kathmandu University
CRISPR/Cas-based gene editing in Daphnia system
I worked on establishing CRISPR/Cas9 and Cas12a-based gene editing techniques for D. pulex and D. magna using microinjection as a ribonucleoprotein (RNP) and plasmid delivery tool. My work was successful in disrupting the gene (knock-out) responsible for eye pigmentation (Scarlet) to generate a white-eyed Daphnia as a validation of the technique.
At present, I am working to introduce a desired nucleotide sequence (knock-in) in a Daphnia gene using the same delivery technique. With positive outcomes so far, my further attempt is to rigorously prepare mutants for other functional genes, like Doublesex (sex-determining gene), CDC20, and insulin receptor genes, to generate a CRISPR mutant library for the Daphnia genome.