Table 2.
Vector systems and inserted target gene 3′-UTR sequences used in reporter gene assays for target gene evaluation of hsa-miR-212-3p and hsa-miR-34c-5p.
| Target gene |
mRNA Acc. number |
Position of 3′-UTR sequence of insert |
miRNA predicted to bind to 3′-UTR sequence |
Location of miRNA binding site in 3′-UTR sequence |
Name and sequence of forward primer |
Name and sequence of reverse primer |
Name of vector system |
Name of vector construct |
|---|---|---|---|---|---|---|---|---|
| ABCG2 | NM_004827.2 | 484–663 | hsa-miR-212-3p | 588–607† | FW-ABCG2-V1: 5′-CTCGTTT′AAACAAAAAAAGCCACCGTGATAGAAA | RV-ABCG2-V1: 5′-GAGT* CTAGAGTTCCAGAAATGGTGCAAGAAT | psicheck2S | ABCG2-V1 |
| ABCG2 | NM_004827.2 | 1328–1462 | hsa-miR-212-3p | 1411–1417‡ | FW-ABCG2-V2: 5′-CTCGTTT′AAACCTCTTTGCTCAGCTTCCCTTAAT | RV-ABCG2-V2: 5′-GAGT* CTAGATTAGTGGAAAAGTGGTGAAATCG | psicheck2S | ABCG2-V2 |
| ADCY1 | NM_021116.2 | 1–612 | hsa-miR-212-3p | 268–290† | FW-ADCY1-V1 5′-CTCGTTT′AAACTGGAGCCCACGTGGGCCTCTGGGGTGCACA |
RV-ADCY1-V1 5′-GAGT* CTAGATTCTCCCCAGACGGTCCAGTTCAAGAGTGGAC |
psicheck2S | ADCY-V1 |
| ADCY1 | NM_021116.2 | 2853–3622 | hsa-miR-212-3p | 3246–3252‡ | FW-ADCY1-V2 5′-CTCGTTT′AAACGATTCGAAGTGTCACAGAGATGGTGCCGTC |
RV-ADCY1-V2 5′-GAGT* CTAGAAGCATGGCCAAGCAAGGGCTGAGGTGGGTGTG |
psicheck2S | ADCY-V2 |
| SOX11 | NM_003108.3 | 51–523 | hsa-miR-212-3p | 509–515‡ | FW-SOX11-V1: 5′-CTCGTTT′AAACTTGGGAGGAAGTTGTAGTGG | RV-SOX11-V1: 5′-GAGT* CTAGACCCTAGAGTAACAGTTGTGC | psicheck2S | SOX11-V1 |
| SOX11 | NM_003108.3 | 6500–7199 | hsa-miR-212-3p | 6620–6641‡,§, 6954–6973† | FW-SOX11-V2: 5′-CTCGTTT′AAACCTCCCTTGAATCACATAGGG | RV-SOX11-V2: 5′-GAGT* CTAGAGAAGCTGGTTAGATCGAAGC | psicheck2S | SOX11-V2 |
| BSN | NM_003458.3 | 362–616 | hsa-miR-212-3p | 485–505† | FW-BSN V1: 5′-CTCGTTT′AAACTACTTCCCACAGCCGCCTTTT | RV-BSN V1: 5′-GAGT* CTAGAGGCCACATGGAGGTCTGCT | psicheck2S | BSN-V1 |
| BSN | NM_003458.3 | 1240–1537 | hsa-miR-212-3p | 1382–1392‡ | FW-BSN V2: 5′-CTCGTTT′AAACGGGGAATGTGGCACTGTCT | RV-BSN V2 5′-GAGT* CTAGA TGCCACAGACTGAGAATTTCAC |
psicheck2S | BSN-V2 |
| MECP2 | NM_004992.3 | 6294–7203 | hsa-miR-212-3p | 6591–6610†, 6887–6893‡,& | FW-MECP2: 5′-CTCGTTT′AAACTCCAGAAACTTGTTCTTCCAAAGC | RV-MECP2: 5′-GAGT* CTAGATACTTTAAACATAGTTACAATACAGA | psicheck2S | MECP2 |
| GABRA3 | NM_000808.3 | 1–1066 | hsa-miR-34c-5p | 221–227‡,§, 256–282†, 722–728‡,§ | FW-GABRA3: 5′-CTCGTTT′AAACATAGTGGTGGCAGTGCAGCAACCAGAGCACTGT | RV-GABRA3: 5′-GAGC# TCGAGTGCCTGGGCCAGGGTGTGGAATGTGGAGTGTA | pmiRGLO | GABRA3 |
| GABBR2 | NM_005458.7 | 259–425 | hsa-miR-34c-5p | 393–418† | FW-GABBR2 V1: 5′-CTCGTTT′AAACAAACAAGGAGGCGCTGGGATAT | RV-GABBR2 V1: 5′-GAGT* CTAGATGTGCAGAGGCAGTTTCTTCAGA | pmiRGLO | GABBR2-V1 |
| GABBR2 | NM_005458.7 | 2282–2433 | hsa-miR-34c-5p | 2384–2389‡ | FW-GABBR2 V2: 5′-CTCGTTT′AAACTCTTTTATATGCAGCAAACACACC | RV-GABBR2 V2: 5′-GAGT* CTAGACATGCAGTAGGGGTACGAGAAATT | pmiRGLO | GABBR2-V2 |
| GRM7 | NM_181874.2 | 1–1196 | hsa-miR-34c-5p | 281–287‡,§, 266–287† | FW-GRM7: 5′-CTCGTTT′AAACCTTTTGACTGCTTTCCCAAAGGCCCTGCTGCA | RV-GRM7: 5′-GAGT* CTAGAAAAAAGTTAACCAAAGTATTTTATTGATACAT | pmiRGLO | GRM7 |
| MBD6 | NM_052897 | 365–869 | hsa-miR-34c-5p | 663–673‡,§, 756–789† | FW-MBD6: 5′-CTCGTTT′AAAC CCCTTCTTAAAGCCAACAGGC | RV-MBD6: 5′-GAGT* CTAGAATAGAAAAGCAGTGGAGGCCC | pmiRGLO | MBD6 |
′ Pme I restriction site
*
Xba I restriction site
#
Xho I restriction site
Predicted by target gene prediction database
†
RNAHybrid,
‡
Targetscan 5.2,
§
miRanda,
&
PicTar