4D Ex Vivo Crispr/Cas9 Whole-Genome Screen to Identify Genes Regulating Lung Cancer Metastasis

<p>Metastatic lung cancer has a 5-year survival rate of 5%. Lung cancers tend to be asymptomatic until late stages, and almost 90% are not diagnosed until they are advanced. Metastases are very rare events, often initiated by a single cell from a primary tumor into a new niche at a distant location. Investigation of the early metastatic process is of urgent need for the development of early diagnostics and targeted therapeutics.<strong> </strong>We performed a proof-of-concept CRISPR/Cas9 whole genome knockout screen in the A549 lung adenocarcinoma cell line and utilized a novel <em>ex vivo 4D </em>lung metastasis model to find gene candidates to test <em>in vitro. </em>I prepared samples and performed next generation sequencing. I created a computational pipeline to demultiplex samples and align reads to the library. I recovered genes previously implicated in lung cancer and metastasis enriched in the Primary and Metastasis samples. SPI1 was identified as the central node in network analysis of the Primary and Metastases and present in every sample. SPI1 was validated <em>in vitro</em> as a novel transcription factor involved in lung cancer metastasis.</p>

Name thesis:degree_name

Doctor of Philosophy (PhD)

Level thesis:degree_level

Dissertation (PhD)

Year dc:date.available

2021

Authors dc:creator

• Plumer, Alexandria
• <p><a href="https://orcid.org/0000-0002-7889-7378">0000-0002-7889-7378</a></p>

Contributors dc:contributor

• Prahlad Ram, Ph.D.
• Faye Johnson, M.D., Ph.D.
• Ju-Seog Lee, Ph.D.

• Metastasis
• Lung Cancer
• SPI1
• CRISPR Screen
• 4D Lung Model
• Cancer Biology
• Cell Biology
• Disease Modeling
• Laboratory and Basic Science Research
• Medicine and Health Sciences

Repository record dc:identifier

https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1146

OAI identifier oai:identifier

oai:digitalcommons.library.tmc.edu:utgsbs_dissertations-2202