Date of Award
Bernie Wone, Ph. D.
Beate Wone, M. S.
Scott Breuninger, Ph. D.
promoter, plasmid, endonuclease
Within studies of plant genome modification there is a method of gene alteration which involves using CRISPR/Cas systems in order to target specific gene loci and implement the intended modification. This process is driven by the ability of a single guide RNA (sgRNA) to find this target location within the plant’s genome. This sgRNA strand is expressed due to the presence of a proper RNA promoter. In an effort to understand the effect of an endogenous RNA promoter’s sequence on the successful expression of a sgRNA strand, an experiment was developed involving twelve U6 promoter sequences from a crassulacean metabolism acid (CAM) plant, Kalanchoe laxiflora. This study was designed to test each promoter sequence individually to determine if it would successfully express the guide RNA (gRNA) strand within a construct. This success would have been verified through the transient expression of the sgRNA/Cas-GFP fusion construct using confocal microscopy.
Jones, Marlee E., "Testing Kalanchoe laxiflora U6 Promoters for Optimal sgRNA Expression in Tobacco" (2020). Honors Thesis. 97.