Julia Hettleman 

The CRISPR-Cas gene editing system is a revolutionary, Nobel prize-awarded discovery that harnesses a natural bacterial defense system to locate and change specific genetic sequences in living cells. This technique has incredible therapeutic potential, and it also has a wide variety of applications that are currently being explored. Researchers in Dr. Harris Wang’s laboratory at Columbia University investigated one of these applications: using CRISPR to activate genes, which means helping genes produce more of the protein they encode. Although CRISPR’s original function is to edit the genome, it can also be used to execute other functions by taking advantage of its effective targeting system. The Wang laboratory was not the first to investigate this type of CRISPR editing, coined CRISPRa, but they wanted to continue perfecting a method upregulating gene expression that was also easily programmable, could be used in various bacterial species, and did not require permanent alterations to the genome. To do this, the researchers wanted to use transcriptional activators, which are proteins that help with the process of copying DNA. Researchers first designed a system that would use a protein called Cas9’s DNA binding properties to bind a transcriptional activator to a predetermined sequence of DNA. They then employed this system in E. Coli cells, testing several different possible transcriptional activators and measuring the efficacy of each one. After performing directed evolution to determine the most effective transcriptional activator variant and repeating the experiment in various species of bacteria, the researchers synthesized a system called dCas9-AsiA-m2.1 that successfully met all their goals for the experiment. This research provides valuable insights into the field of gene editing and this CRISPRa system, as well as the various research techniques used in this study, can surely facilitate future research.