Table 1. DNA oligos for siRNA production.
| Target Site | DNA templates |
|---|---|
| 437–4611 | Oligo1: GGATCCTAATACGACTCACTATAGCAAGCTCCTGAAGTGGGAAGTGAA2 |
| 437–4611 | Oligo2:AATTCACTTCCCACTTCAGGAGCTTGCTATAGTGAGTCGTATTAGGATCC2 |
| 437–4611 | Oligo3: GGATCCTAATACGACTCACTATATTCACTTCCCACTTCAGGAGCTTGC2 |
| 437–4611 | Oligo4:AAGCAAGCTCCTGAAGTGGGAAGTGAATATAGTGAGTCGTATTAGGATCC2 |
| 631–6341 | Oligo1: GGATCCTAATACGACTCACTATAGAGAAGAGACTGCTTGATAACATA2 |
| 631–6341 | Oligo2:AATATGTTATCAAGCAGTCTCTTCTCTATAGTGAGTCGTATTAGGATCC2 |
| 631–6341 | Oligo3: GGATCCTAATACGACTCACTATATATGTTATCAAGCAGTCTCTTCTC2 |
| 631–6341 | Oligo4:AAGAGAAGAGACTGCTTGATAACATATATAGTGAGTCGTATTAGGATCC2 |
| 718–7421 | Oligo1: GGATCCTAATACGACTCACTATAGAGCTCTCCGAAGCCAAGATGTTGT2 |
| 718–7421 | Oligo2:AAACAACATCTTGGCTTCGGAGAGCTCTATAGTGAGTCGTATTAGGATCC2 |
| 718–7421 | Oligo3: GGATCCTAATACGACTCACTATAACAACATCTTGGCTTCGGAGAGCTC2 |
| 718–7421 | Oligo4:AAGAGCTCTCCGAAGCCAAGATGTTGTTATAGTGAGTCGTATTAGGATCC2 |
| 880–9021 | Oligo1: GGATCCTAATACGACTCACTATAGACAACGTCACTCTAGACCTGAA2 |
| 880–9021 | Oligo2:AATTCAGGTCTAGAGTGACGTTGTCTATAGTGAGTCGTATTAGGATCC2 |
| 880–9021 | Oligo3: GGATCCTAATACGACTCACTATATTCAGGTCTAGAGTGACGTTGTC2 |
| 880–9021 | Oligo4:AAGACAACGTCACTCTAGACCTGAATATAGTGAGTCGTATTAGGATCC2 |
| Control3 | Oligo1: GGATCCTAATACGACTCACTATACCTACGCCACCAATTTCGTTT2 |
| Control3 | Oligo2:AAAAACGAAATTGGTGGCGTAGGTATAGTGAGTCGTATTAGGATCC2 |
| Control3 | Oligo3: GGATCCTAATACGACTCACTATAAAACGAAATTGGTGGCGTAGG2 |
| Control3 | Oligo4:AACCTACGCCACCAATTTCGTTTTATAGTGAGTCGTATTAGGATCC2 |
1 The numbers are the starting and ending position of the target sites in TRIM75 CDS (NM_001033429.2 in NCBI).
2 two pairs of DNA oligos are needed for for each double-stand siRNA. Oligo 2 is complementary with oligo 1 except an "AA" overhang at 5'; Oligo 3 is complementary with oligo 4 except an "AA" overhang at 5'. Anealed Oligo 1 and 2 will be the template for the forward stand of siRNA and anealed Oligo 3 and 4 will be the template for the reverse complementary strand of siRNA. Reactions for the production of forward or reverse complementary single-stand siRNA are set up separately, finally combined together and annealed to generate a stable, ready-to-use double-strand siRNA. In each oligo, gene-specific sequences are underlined, other sequences are for recognition and binding by T7 RNA polymerase.
3 Control siRNA does not target to any mRNA sequence in mouse.