miR-217 mimics, scramble mimics, anti-miR-NC (negative control), and anti-miR-217 were synthesized by GenePharma (Shanghai, People’s Republic of China). miRNAs (10 nM) were transfected into cervical cancer cell using DharmaFECT1 Reagent (Dharmacon, Lafayette, CO, USA). To construct stably overexpressing miR-217 SiHa cell, lentiviral constructs (LentimiR™ miRNA precursor clones; System Biosciences, Palo Alto, CA, USA) expressing miR-217 or respective empty vector (miR-NC) were packaged using the pPACKH1 Lentivector Packaging System (System Biosciences) and were used to transfect into SiHa cell. Stable clones were selected using 1 µg/mL puromycin (Thermo Fisher Scientific).20 (link),21 (link)
Protocol detail
Transfection and Stable Overexpression of miR-217 in Cervical Cancer Cells
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Variable analysis
independent variables
- MiR-217 mimics
- Scramble mimics
- Anti-miR-NC (negative control)
- Anti-miR-217
- Stable overexpression of miR-217 in SiHa cell
- Outcomes or effects in cervical cancer cells
- DharmaFECT1 Reagent used for transfection
- Puromycin (1 µg/mL) used for selecting stable clones
- Empty vector (miR-NC) used as control for stable overexpression
- Negative control: anti-miR-NC
- Positive control: Not explicitly mentioned
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