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- Title
Knockout of <italic>CTNNB1</italic> by CRISPR-Cas9 technology inhibits cell proliferation through the Wnt/β-catenin signaling pathway.
- Authors
Guan, Lihong; Zhu, Shaoyi; Han, Yawei; Yang, Ciqing; Liu, Yanli; Qiao, Liang; Li, Xiaoying; Li, Han; Lin, Juntang
- Abstract
Objective: To study the effects of <italic>CTNNB1</italic> gene knockout by CRISPR-Cas9 technology on cell adhesion, proliferation, apoptosis, and Wnt/β-catenin signaling pathway.Results: <italic>CTNNB1</italic> gene of HEK 293T cells was knocked out by CRISPR-Cas9. This was confirmed by sequencing and western blotting. Methylthiazolyl-tetrazolium bromide assays indicated that deletion of β-catenin significantly weakened adhesion ability and inhibited proliferation rate (<italic>P</italic> < 0.01) of HEK 293T cells. Nevertheless, deletion of β-catenin did not affect apoptosis of HEK 293T cells, which was analyzed by flow cytometry with Annexin V-fluorescein isothiocyanate/propidium iodide double staining. In addition, expression level of <italic>GSK</italic>-<italic>3β</italic>, <italic>CCND1</italic>, and <italic>CCNE1</italic> detected by qPCR and expression level of N-Cadherin and cyclin D1 detected by western blotting were significantly decreased (<italic>P</italic> < 0.01) while expression of γ-catenin detected by western blotting was significantly increased (<italic>P</italic> < 0.001).Conclusions: Knockout of <italic>CTNNB1</italic> disturbed Wnt/β-catenin signaling pathway and significantly inhibited adhesion and proliferation of HEK 293T cells.
- Subjects
CRISPRS; CELL proliferation; CATENINS; CELL communication; WNT genes; APOPTOSIS
- Publication
Biotechnology Letters, 2018, Vol 40, Issue 3, p501
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-017-2491-2