A CRISPR platform for rapid and inducible genome editing in human non-small cell lung cancer cells
Grade Level at Time of Presentation
Senior
Major
Cellular/Physiology
Minor
n/a
Institution
University of Louisville
KY House District #
40
KY Senate District #
35
Faculty Advisor/ Mentor
Dr. Chi Li
Department
University of Louisville School of Medicine, Pharmacology & Toxicology
Abstract
Non-small cell lung cancer (NSCLC) accounts for about 85% of lung cancer, which is the leading cause of cancer death in the world. High mortality rate associated with NSCLC is partially attributed to the limited understanding of NSCLC as well as ineffective therapeutic treatments. The initiation and progression of NSCLC involves genetic changes leading to alterations in the control of tissue development and homeostatic maintenance. Better knowledge about these genetic abnormalities is imperative for developing new chemotherapeutic drugs for NSCLC. Recent research demonstrates that the expression of paraoxonase 2 (PON2), a lactonase/arylesterase with anti-oxidant properties, are markedly enhanced in cancer tissues of NSCLC patients compared with corresponding adjacent non-tumorous tissues. Importantly, increased PON2 expression likely contributes to the resistance of NSCLC cells to classical anti-NSCLC therapeutic drugs. We found that stably reduced PON2 expression by siRNA reduced the proliferation of NSCLC cells but not their untransformed epithelia counterparts. To further elucidate the role of PON2 in NSCLC cell proliferation, we applied two newly developed gene-editing systems, TALEN and CRISPR/Cas, in the NSCLC cell line NCI-H1299. Our data indicated that Cas9 expression was induced by exogenous doxycycline in NSCLC cells in a reversible fashion. The goal of this study is to establish an efficient and scalable experimental approach interrupting the expression of a gene (e.g. PON2) in NSCLC cells, which provides a platform to investigate how changes in gene expression modulate NSCLC cell proliferation.
A CRISPR platform for rapid and inducible genome editing in human non-small cell lung cancer cells
Non-small cell lung cancer (NSCLC) accounts for about 85% of lung cancer, which is the leading cause of cancer death in the world. High mortality rate associated with NSCLC is partially attributed to the limited understanding of NSCLC as well as ineffective therapeutic treatments. The initiation and progression of NSCLC involves genetic changes leading to alterations in the control of tissue development and homeostatic maintenance. Better knowledge about these genetic abnormalities is imperative for developing new chemotherapeutic drugs for NSCLC. Recent research demonstrates that the expression of paraoxonase 2 (PON2), a lactonase/arylesterase with anti-oxidant properties, are markedly enhanced in cancer tissues of NSCLC patients compared with corresponding adjacent non-tumorous tissues. Importantly, increased PON2 expression likely contributes to the resistance of NSCLC cells to classical anti-NSCLC therapeutic drugs. We found that stably reduced PON2 expression by siRNA reduced the proliferation of NSCLC cells but not their untransformed epithelia counterparts. To further elucidate the role of PON2 in NSCLC cell proliferation, we applied two newly developed gene-editing systems, TALEN and CRISPR/Cas, in the NSCLC cell line NCI-H1299. Our data indicated that Cas9 expression was induced by exogenous doxycycline in NSCLC cells in a reversible fashion. The goal of this study is to establish an efficient and scalable experimental approach interrupting the expression of a gene (e.g. PON2) in NSCLC cells, which provides a platform to investigate how changes in gene expression modulate NSCLC cell proliferation.