CRISPR-Cas9 as a gene editing tool using cardiac glycoside reductase operon for digoxin metabolism

Gene editing technology has gained popularity over the past two decades with the scientific advancements being made in genetics and computational biology. CRISPR-Cas9 technique allows us to target genetic material and perform accurate manipulation of targeted gene function. Cgr operon, commonly found in the gut microbiome bacteria of E. lenta has several attributes which can be modified using the CRISPR-Cas9 methodology. Here, we briefly describe how the Cas9 nuclease can be used to prevent Digoxin from being suppressed by the cgr gene, directly correlating to improved cardiac function. The editing tool is described to assist in mutating the genetic material of cgr gene discussing the inhibition of Na+/K+ ATPase in cardiac myocytes and CG-content. The principle of operation behind CRISPR-Cas9 is described using non-homologous end joining or homology-directed repair cellular mechanisms. Target site selection, designing sgRNAs and introducing various mutations are discussed with limitations of current technology and possible applications in the field of point of care diagnostics and biosensing developing therapeutic interventions using cgr operon.