Date of Award

Spring 5-2020

Document Type

Honors Thesis

Degree Name

Bachelor of Science

Department

Biology

Advisor/Committee Chair

Arun Richard Chandrasekaran, Ph.D.

Major

Biology

Committee Member

Ken Halvorsen, Ph.D.

Committee Member

Elise Gervais, Ph.D.

Abstract

Ribonuclease H (RNase H) is a damage-repair protein and ribonuclease that specifically catalyzes the hydrolysis of RNA in an RNA/DNA duplex and breaks down RNA/DNA junctions. It plays an important role in a variety of biological processes including DNA replication, DNA repair, and transcription. It is also pivotal in anti-HIV drug development and the analysis of cellular processes and has been shown to be a potential therapeutic target for various neoplastic diseases. This thesis discusses a unique assay based on DNA nanoswitches to detect RNase H levels and activity. The assay is based on conformational changes of DNA nanoswitches in the presence of RNase H. Using gel electrophoresis, we study the kinetics of cleavage, sensitivity and specificity of detection of RNase H. We also show the utility of the assay in screening potential RNase H inhibitors. This novel biosensing platform is low-cost, label-free, and will be a useful tool to detect RNase levels and identify RNase contamination in biological samples.

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