I chose to collect data using Existing Data Analysis because there is already an

abundance of data and studies that I can use to support my thesis without without conducting an

experiment. If I were to conduct an experiment, I would be repeating the research of other

scientists, and it would be a waste of time since I do not have the resources and time to do any

extensive experimental research needed to effectively complete a study. My data applies to my

thesis by supporting the thesis with legitimate information backed up by controlled and well-

organized experiments conducted by professionals. The data from the studies that I used

supported the part of my thesis that was relevant to new, innovative methods to improve the on-

target activity of CRISPR-Cas9. Many of these methods involve using a different version of

Cas9, the protein that cuts the DNA. These versions of Cas9, called SpCas9, have higher editing

rates and are shown to greatly increase the specificity of Cas9. Furthermore, lab created

variations of the wild-type SpCas9 can improve the specificity of Cas9 so much as off-target

edits are undetected when using these variations. This information effectively supports my thesis

because it describes innovative ways to improve the on-target activity of CRISPR-Cas9 in order

to make it safer for gene therapy treatments. My results showed the SpCas9 was the most

effective method to improving the specificity because the data showed that compared to Cas9,

SpCas9 was able to reduce off-target edits to the point where they were undetectable. The studies

that I gathered data from also showed that certain variations of the wild-type SpCas9 were able

to increase specificity even more in human genes. As well as improving specificity itself, some

sources also described using computerized programs to detect any off-target effects that may

have occurred. This also relates to my thesis because it is an innovative way to make sure that

CRISPR is safe.