A newly engineered CRISPR protein could help scientists observe the molecular scissors called Cas9 enzyme as it enables them to edit genomes using the CRISPR-Cas9 system for treating genetic diseases including cancer.
Gene therapy could be a permanent cure for many life-threatening hereditary diseases. Developing effective, affordable, and safe gene therapy methods remained a challenge for decades. A major breakthrough to address this challenge came in the form of CRISPR, a gene-editing tool that uses a guide RNA to direct a Cas9 enzyme to a specific DNA sequence, where it precisely cuts the DNA. The CRISPR-Cas9 had been designed to cut and correct DNA with accuracy. However, scientists could not observe Cas9, the molecular surgeon, in living cells in real time. Traditional detection methods rely on fixing or breaking open cells, making it impossible to track the process as it unfolds.
Tracking gene editing as it happens or watching the molecular machinery as it works, cutting, repairing, and rewriting DNA inside living cells can help monitor CRISPR operations in living cells and tissues without destroying them.
Scientists from Kolkata based Bose institute, an autonomous institute of the Department of Science and Technology (DST), has come out with a solution for this. A team led by Dr. Basudeb Maji, has created GlowCas9, a CRISPR protein that lights up while performing gene editing. Their study, published in Angewandte Chemie International Edition opens a new chapter in the visualization and tracking of genome engineering.
Arkadeep Karmakar, a Ph.D. researcher in Dr. Basudeb Maji’s lab, designed GlowCas9, a bioluminescent version of Cas9 that glows inside cells, by fusing Cas9 with a split nano-luciferase enzyme derived from deep-sea shrimp proteins.