Establishment of an miR‑137‑knockout cell model using CRISPR/Cas9 genome editing

  • Authors:
    • Wei Chen
    • Qi Li
    • Jingjie Du
    • Xiaodi Li
    • Songshan Jiang
    • Yuanli He
  • View Affiliations

  • Published online on: July 6, 2018     https://doi.org/10.3892/ol.2018.9096
  • Pages: 4027-4032
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Abstract

MicroRNA‑137 (miR‑137) has been reported to be abnormally expressed in a variety of types of cancer, including ovarian cancer. However, the roles served by miR‑137 in cancer are not fully understood. In the present study, 3 single guide RNAs (sgRNAs) were designed, synthesized and inserted into pXPR001 plasmids. The pXPR001‑sgRNA plasmids were verified using sequencing and integrated into the genome of the ovarian cancer cell line, A2780, through lentiviral transduction, puromycin selection and single‑cell culture. PCR products amplified from single‑cell cultures using primers covering miR‑137 targeting sites were sequenced to identify clones with miR‑137 gene disruption. Genome editing was detected in 72% of the clones derived from sgRNA2, 4% from sgRNA3 and 0% from sgRNA1. Of the clones from sgRNA2, 32% contained 1 edited miR‑137 allele and 40% contained 2 edited miR‑137 alleles. The expression of miR‑137 in clones #2 and #3 could not be detected by reverse transcription‑­quantitative polymerase chain reaction. In addition, an MTT assay demonstrated that clones #2 and #3 exhibited enhanced proliferation. In conclusion, an miR‑137‑knockout cell model was successfully established in A2780 cells using CRISPR/Cas9 technology.
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September-2018
Volume 16 Issue 3

Print ISSN: 1792-1074
Online ISSN:1792-1082

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Copy and paste a formatted citation
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Spandidos Publications style
Chen W, Li Q, Du J, Li X, Jiang S and He Y: Establishment of an miR‑137‑knockout cell model using CRISPR/Cas9 genome editing. Oncol Lett 16: 4027-4032, 2018
APA
Chen, W., Li, Q., Du, J., Li, X., Jiang, S., & He, Y. (2018). Establishment of an miR‑137‑knockout cell model using CRISPR/Cas9 genome editing. Oncology Letters, 16, 4027-4032. https://doi.org/10.3892/ol.2018.9096
MLA
Chen, W., Li, Q., Du, J., Li, X., Jiang, S., He, Y."Establishment of an miR‑137‑knockout cell model using CRISPR/Cas9 genome editing". Oncology Letters 16.3 (2018): 4027-4032.
Chicago
Chen, W., Li, Q., Du, J., Li, X., Jiang, S., He, Y."Establishment of an miR‑137‑knockout cell model using CRISPR/Cas9 genome editing". Oncology Letters 16, no. 3 (2018): 4027-4032. https://doi.org/10.3892/ol.2018.9096