The 3′UTR fragments of CCNG1 containing putative binding sites for miR-1271 were cloned into pMIR-Report construct (Ambion, Austin, TX). The primers (Biomart, Shanghai, China) were constructed according to previous reports [33 (link),34 (link)]. Mutant 3′UTR of CCNG1, which carried a mutated sequence in the complementary site for the seed region of miR-1271, were generated using the fusion PCR method. Luciferase reporter assay was performed in HEK293 cells as described previously [12 (link)].
Protocol detail
Luciferase Reporter Assay for miR-1271 Targeting
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Corresponding Organization :
Other organizations : Sixth Affiliated Hospital of Kunming Medical University, Kunming Medical University
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Variable analysis
independent variables
- Mutant 3'UTR of CCNG1, which carried a mutated sequence in the complementary site for the seed region of miR-1271
- Luciferase reporter activity
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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