Deletion of FOXL2 by CRISPR promotes cell cycle G0/G1 restriction in KGN cells

  • Authors:
    • Bin Tang
    • Yujie Zhang
    • Wei Zhang
    • Yuqing Zhu
    • Shaopeng Yuan
  • View Affiliations

  • Published online on: October 24, 2018     https://doi.org/10.3892/ijmm.2018.3956
  • Pages: 567-574
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Abstract

Forkhead box L2 (FOXL2), a member of the forkhead family of transcription factors, is important in eyelid and ovary differentiation. Although the function of FOXL2 in organogenesis has been investigated, the detailed mechanisms by which FOXL2 mediates cellular process remain to be fully elucidated. Few FOXL2‑knockout cell lines have been reported, which has limited molecular mechanism investigations. CRISPR is a novel gene editing technique that has been widely used in human genetic diseases. In the present study, FOXL2 was disrupted using clustered regularly interspaced short palindromic repeats (CRISPR), and screening of a stable knockout cell line was performed in human ovarian granulosa KGN cells. Three sites (F404, F425 and F446) around the ATG start codon on the FOXL2 DNA sequence were constructed in a guide RNA lentivirus. Targeting F425 was most efficient, and western blot analysis and DNA sequencing of the resulting cells suggested that both FOXL2 alleles were fully disrupted. In addition, flow cytometry results indicated that the knockout of FOXL2 restricted cell cycle progression at the G0/G1 phase. In addition, the expression levels of cell cycle mediators cyclin D1 and cyclin‑dependent kinase 4 were reduced. These results confirmed that FOXL2 disruption in KGN cells is associated with the cell cycle attenuation.
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January 2019
Volume 43 Issue 1

Print ISSN: 1107-3756
Online ISSN:1791-244X

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APA
Tang, B., Zhang, Y., Zhang, W., Zhu, Y., & Yuan, S. (2019). Deletion of FOXL2 by CRISPR promotes cell cycle G0/G1 restriction in KGN cells. International Journal of Molecular Medicine, 43, 567-574. https://doi.org/10.3892/ijmm.2018.3956
MLA
Tang, B., Zhang, Y., Zhang, W., Zhu, Y., Yuan, S."Deletion of FOXL2 by CRISPR promotes cell cycle G0/G1 restriction in KGN cells". International Journal of Molecular Medicine 43.1 (2019): 567-574.
Chicago
Tang, B., Zhang, Y., Zhang, W., Zhu, Y., Yuan, S."Deletion of FOXL2 by CRISPR promotes cell cycle G0/G1 restriction in KGN cells". International Journal of Molecular Medicine 43, no. 1 (2019): 567-574. https://doi.org/10.3892/ijmm.2018.3956