CRISPR-Cas9 Mediated Deletion of Glutaredoxin in Lung Epithelial-like Cancer Cells

Presenter's Name(s)

Alexander D'Amico

Conference Year

January 2021

Abstract

Idiopathic pulmonary fibrosis is a deadly respiratory disease accompanied by changes in the redox environment of the lung. Dr. Janssen-Heininger’s lab has shown reversal of lung fibrosis in mice by administering glutaredoxin protein. Based on these findings, the lab is currently focused on determining the mechanism by which glutaredoxin had this effect. We are therefore interested in generating loss-of-function lung epithelial cell lines using CRISPR-Cas9 technology. In order to demonstrate efficient glutaredoxin knockout using CRISPR-Cas9, we first applied this methodology to human lung epithelial-like cancer cells (H522). We screened for glutaredoxin deletion in H522 cell cultures and found multiple colonies lacking glutaredoxin expression. These promising data will now allow us to apply this approach to lung cell lines that are more difficult to culture, such as normal lung epithelia, in an effort to determine the roles of glutaredoxin in lung homeostasis and disease.

Primary Faculty Mentor Name

Yvonne Janssen-Heininger, Ph.D.

Secondary Mentor Name

Joseph Druso, Ph.D.

Faculty/Staff Collaborators

Zhihua Peng, Reem Aboushousha

Status

Undergraduate

Student College

College of Arts and Sciences

Program/Major

Biological Science

Primary Research Category

Biological Sciences

Secondary Research Category

Health Sciences

Abstract only.

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CRISPR-Cas9 Mediated Deletion of Glutaredoxin in Lung Epithelial-like Cancer Cells

Idiopathic pulmonary fibrosis is a deadly respiratory disease accompanied by changes in the redox environment of the lung. Dr. Janssen-Heininger’s lab has shown reversal of lung fibrosis in mice by administering glutaredoxin protein. Based on these findings, the lab is currently focused on determining the mechanism by which glutaredoxin had this effect. We are therefore interested in generating loss-of-function lung epithelial cell lines using CRISPR-Cas9 technology. In order to demonstrate efficient glutaredoxin knockout using CRISPR-Cas9, we first applied this methodology to human lung epithelial-like cancer cells (H522). We screened for glutaredoxin deletion in H522 cell cultures and found multiple colonies lacking glutaredoxin expression. These promising data will now allow us to apply this approach to lung cell lines that are more difficult to culture, such as normal lung epithelia, in an effort to determine the roles of glutaredoxin in lung homeostasis and disease.