Effect of Gene Knockout of CEP290 using CRISPR-Cas 9 Technology on Xenopus tropicalis

Abstract

Modelling human diseases using the Xenopus species is an increasingly popular method to study vertebrate embryology and development, basic cell and molecular biology, genomics, neurobiology, and toxicology. This allows for the elucidation of the regulation mechanisms and interactive networks that affect the direct development of embryos, the adaptation process, and disease and malformation-causing dysregulations. Here we aim to analyze the possible kidney defects during the gene-knockout of CEP290 in X. tropicalis species. Our objectives are to produce sgRNA via de-novo synthesis from the constructed DNA template, to microinject synthesized sgRNA into embryos of Xenopus tropicalis, and to determin the success of the genome editing via T7-Endonuclease I assay and observation of genetically modified tadpoles for identifying any physical symptoms produced due to gene knockout. The data obtained from the embryos of X. tropicalis suggests that complete knock-out of the gene cep290 results in severe mutation that causes death.