Date of Award

5-2017

Document Type

Honors Thesis

Degree Name

Bachelor of Science

Department

Nanoscale Science

First Advisor

Thomas Begley

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR) is a precise genetic engineering tool for genome editing. CRISPR utilizes guide RNA (gRNA) to find specific DNA sequences followed by a Cas9 nuclease to cut the DNA at a specific site. TRM9L is a tumor growth-suppresser gene that restricts the growth of some colorectal cancer cells by upregulating LIN9 expression. TRM9L expression is lost in some late stage colorectal cancers and cancer models (SW620). SW480 colorectal cancer cells express TRM9L and these cells are considered to be at the beginning of colorectal cancer development. The goal of my project was to use genetic engineering approaches to remove TRM9L from early stage colorectal cancer cells (SW480) and to then determine the effect on tumor growth. In this project, CRISPR constructs were synthesized, hybridized, and ligated to an expression vector. The hybridized vector underwent bacterial-facilitated replication and was then harvested via plasmid purification. The plasmids were gene sequenced and then transfected into a growing culture of SW480 cells. After a month of plasmid-inclusion growth, the cell cultures will be analyzed for growth effects. My work is important because it will develop new tools for use in colorectal cancer research, a disease that claims over 50,000 lives each year.

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