A CRISPR way of editing genes (still working on the crispness though...)

Genetic engineering is something that has long fascinated us. It holds the secret to eradicating genetic disorders, studying the function of almost anything in biological systems, and creating “perfect” organisms (where organism is a loose term that could refer to anything from plants in agricultural practice to humans in the 1997 movie Gattaca). So of course, the development of CRISPR created quite a stir the world over. And with good reason, I think. 

It’s still quite unbelievable that in Mod 2 we were able to sit at a lab bench, order some oligos from a company, mix some things together (alright, this may be a tad oversimplified) and cause a lasting change in E. Coli. A change they pass down to their daughter cells, which those daughter cells will pass to their daughter cells and then their daughter… Well anyway, the point is that we are doing some amazing, cutting-edge stuff. 

At my UROP, I’ve been working on using CRISPR to mediate homology-directed repair to convert GFP in HeLa cells to BFP, effectively allowing the cells to express blue fluorescence rather than green. The first time we visualized our transfected cells, we disappointedly scrolled through the sea of green in our plate until we saw EXACTLY ONE bright blue spot. Yes, it seems we had exactly one cell do what we wanted our hundreds of thousands of cells to do. Either way, it was beyond exciting to see that it actually worked. So when I analyzed the ethanol production in our CRISPR-modified (and very stinky) E. Coli in Mod 2, I was awed that our experimental samples had a SEVERAL fold increase in ethanol production over the control ones. 

Truthfully, I was pretty skeptical that our experiments would work. SO many things could have gone wrong: bad oligo design, ineffective plasmid transformation, dCas9 not preventing transcription well enough. And I mean, there’s always something that goes mysteriously wrong. But our experiments, for the most part, worked! While there are definitely some kinks that need to be worked out of the CRISPR-Cas9 system, the possibilities this technology opens up are incredibly exciting.