Citlalli Grijalva

The evolving technology of gene editing

During the two weeks at the Fred Hutch Explorers program a highlight of mine would be the different lab procedures I got to do, in particular the CRISPR wet lab in which we practiced the CRISPR method. What the CRISPR method specifically does is it uses an enzyme called Cas9 to cut a piece of DNA from a specific gene but what makes it truly remarkable is that one can program it so that the cut is specific to their needs. In our wet lab we had the opportunity to model how the CRISPR method worked and how it could be applied to real life situations. However, in order to practice the CRISPR technique in our lab we first had to learn about the many steps that go into it. For example, we first had to learn how to replicate multiple copies of a section of a BRCA1 gene so that we could sufficient DNA to successfully cut the DNA using the CRISPR method. We specifically chose the BRCA1 gene as it’s known to have a mutation in the DNA that could cause to breast cancer. Once that step was completed we took our product and tested it using gel electrophoresis to make sure we had made enough copies of our section of DNA.When we knew we had sufficient DNA we then mixed it with our Cas9 enzyme and our guide RNAs that led the enzyme to where we wanted our DNA to be exactly cut. After we had left our product to incubate and added the remaining chemicals we used gel electrophoresis again to see where the Cas9 made the cuts in the DNA and if it was accurate. While we only got to see how the CRISPR method works there are many real world applications that are undergoing using this method. Since it was only just discovered recently scientists are still figuring how they can use this method to treat diseases that are caused by genetic mutations.

What I found the most intriguing about this specific lab was the newfound simplicity of a gene editing technique that was considered more efficient than other techniques. In addition, there are research currently developing to see if they can treat genetic diseases, for example, muscular dystrophy. This particular experience made an impression on me as this lab was very different but also very relevant to the work scientists are currently doing today. The CRISPR method in general was very fascinating as it’s developing science which goes to show the many progressions in science but also the need of improvements of already existing methods.

In the left picture you can see how the gel electrophoresis was set up so that we could see our results from our CRISPR experiment.

In the above picture you can see that we were in the process of making our PCR product so that we had sufficient DNA to conduct our experiment

In the picture above is our results from the gel electrophoresis to check if we had enough DNA in our PCR product to conduct our experiment. We determined the amount by using the DNA ladder to check how many base pairs were in the product.