Date of Award
Master of Science (MS)
Jeffry L. Shultz
Restriction digests are a commonly utilized process for cleaving DNA at specific, but relatively common sites. Restriction enzymes have widespread use in DNA manipulation. CRISPR/Cas9 is a recently identified endonuclease which utilizes a customizable guide sequence to recognize and cut specific ~20 bp sites located in a DNA sequence. This preliminary research aimed to exploit the potential benefit of DNA restriction using the CRISPR/Cas9 procedure through alterations of different components involved in that system. We sought to refine existing CRISPR/Cas9 protocols and make a budget friendly, user-selectable CRISPR/Cas9 restriction digest protocol. The motivation for this research was to simplify and adapt known CRISPR protocols in hopes of using CRISPR as a targeted restriction enzyme. This project yielded negative results, however, important insights into the dilution of target sequences was achieved.
Crawley, Zack, "" (2020). Thesis. 32.