Designing a Novel CRISPR/Cas9 Endogenous Tag of the Amph-1 Gene for Treatment Research Purposes

Abstract

Amphiphysin 1, or amph-1, is a c. elegans ortholog of the genes amphiphysin (AMPH) and bridging integrator 1 (BIN 1) present in humans. Amph-1 is located primarily in the endomembrane system of the cell. Its protein count is highly affected by diseases under dementia and carcinoma. To gain further understanding of amph-1 protein activity in response to such diseases, this research centers around the creation of a CRISPR/Cas9 design plan to inject modified plasmids into c. elegans eggs and produce specimens available for endogenous activity research. An N20 sequence and 5’ and 3’ homology arms were located near the N-terminus, where a bioluminescent protein tag would be inserted. A plasmid dedicated to synthesizing the Cas9 protein and a second plasmid carrying the genetic code for bioluminescent protein are then modified using amph-1 genetic code as a base template. Following silent mutation edits to the N20 sequence, this plasmid design has potential to be developed in labs for depper research into the amph-1 gene behavior.