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.
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