Date of Award

Spring 5-9-2020

Document Type

Honors Thesis



First Advisor

Bernie Wone, Ph. D.

Second Advisor

Beate Wone, M. S.

Third Advisor

Scott Breuninger, Ph. D.


promoter, plasmid, endonuclease

Subject Categories

Molecular Biology


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.



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