CRISPR-Cas9 mediated nox4 knockout inhibits cell proliferation and invasion in hela cells

Naser Jafari, Hyunju Kim, Rackhyun Park, Liqing Li, Minsu Jang, Andrew J. Morris, Junsoo Park, Cai Huang

Research output: Contribution to journalArticle

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Abstract

Increased expression of NOX4 protein is associated with cancer progression and metastasis but the role of NOX4 in cell proliferation and invasion is not fully understood. We generated NOX4 knockout HeLa cell lines using the CRISPR-Cas9 gene editing system to explore the cellular functions of NOX4. After transfection of CRISPR-Cas9 construct, we performed T7 endonuclease 1 assays and DNA sequencing to generate and identify insertion and deletion of the NOX4 locus. We confirmed the knockout of NOX4 by Western blotting. NOX4 knockout cell lines showed reduced cell proliferation with an increase of sub-G1 cell population and the decrease of S/G2/M population. Moreover, NOX4 deficiency resulted in a dramatic decrease in invadopodium formation and the invasive activity. In addition, NOX4 deficiency also caused a decrease in focal adhesions and cell migration in HeLa cells. These results suggest that NOX4 is required for both efficient proliferation and invasion of HeLa cells.

Original languageEnglish
Article numbere0170327
JournalPloS one
Volume12
Issue number1
DOIs
Publication statusPublished - 2017 Jan 1

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Clustered Regularly Interspaced Short Palindromic Repeats
cell invasion
Cell proliferation
HeLa Cells
cell proliferation
Cells
cell lines
Cell Proliferation
transfection
cell movement
metastasis
adhesion
Cell Line
Western blotting
Focal Adhesions
sequence analysis
protein synthesis
Endonucleases
Population Dynamics
cells

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Jafari, N., Kim, H., Park, R., Li, L., Jang, M., Morris, A. J., ... Huang, C. (2017). CRISPR-Cas9 mediated nox4 knockout inhibits cell proliferation and invasion in hela cells. PloS one, 12(1), [e0170327]. https://doi.org/10.1371/journal.pone.0170327
Jafari, Naser ; Kim, Hyunju ; Park, Rackhyun ; Li, Liqing ; Jang, Minsu ; Morris, Andrew J. ; Park, Junsoo ; Huang, Cai. / CRISPR-Cas9 mediated nox4 knockout inhibits cell proliferation and invasion in hela cells. In: PloS one. 2017 ; Vol. 12, No. 1.
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Jafari, N, Kim, H, Park, R, Li, L, Jang, M, Morris, AJ, Park, J & Huang, C 2017, 'CRISPR-Cas9 mediated nox4 knockout inhibits cell proliferation and invasion in hela cells', PloS one, vol. 12, no. 1, e0170327. https://doi.org/10.1371/journal.pone.0170327

CRISPR-Cas9 mediated nox4 knockout inhibits cell proliferation and invasion in hela cells. / Jafari, Naser; Kim, Hyunju; Park, Rackhyun; Li, Liqing; Jang, Minsu; Morris, Andrew J.; Park, Junsoo; Huang, Cai.

In: PloS one, Vol. 12, No. 1, e0170327, 01.01.2017.

Research output: Contribution to journalArticle

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