A polyoligonucleotide molecule and its application in multi-target interference
A technology of polynucleotides and oligonucleotides, which is applied in the field of polyoligonucleotide molecules and their application in multi-target interference, can solve the problem that a single molecule cannot be guaranteed to inhibit target gene expression, endonuclease and exonuclease Highly sensitive enzymes, cumbersome synthesis steps and other issues, to achieve the effect of multi-target interference, RNAi efficiency improvement, and broad application prospects
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Embodiment 1
[0130] Embodiment 1, the design and synthesis of polyoligonucleotide molecule
[0131] 1. Design of Polyoligonucleotide Molecules
[0132] The polyoligonucleotide molecule of the present invention comprises at least 3 oligonucleotide sequences, the size of each oligonucleotide sequence in the polyoligonucleotide molecule is 15-50nt, and each oligonucleotide sequence starts at the 5' end The 3' end is sequentially composed of a targeting terminal fragment TTS with a size of 5-24nt, a targeted internal fragment TIS with a size of 1-20nt, and an adapter terminal fragment ATS with a size of 5-24nt. Each oligonucleotide sequence in the polyoligonucleotide molecule forms a double-stranded region with the other two sequences in the polyoligonucleotide molecule except itself through sequence complementarity, that is, the targeting terminal of each oligonucleotide The fragment TTS is complementary to the adapter terminal fragment ATS of its adjacent oligonucleotide; and the targeting ...
Embodiment 2
[0143] Example 2. Application of Polyoligonucleotide Molecules in Inhibiting Target Gene Expression in Tumor Cells
[0144] (1) Inhibition test for the same target gene
[0145] 1. Molecular inhibition test of polyoligonucleotides with different numbers of sequences
[0146] 1. Molecular inhibition test of polyoligonucleotide for different sequence numbers of p65 gene
[0147] (1) Preparation of polyoligonucleotide molecules
[0148] Three kinds of polyoligonucleotides N(R m1 +R m2 +R m3 ), namely 4(R12+R6+R12), 5(R12+R6+R12), 6(R12+R6+R12). Among them, N represents the number of sequences, m1 represents the nucleotide number of the TTS fragment of each sequence, m2 represents the nucleotide number of the TIS fragment of each sequence, and m3 represents the nucleotide number of the ATS fragment of each sequence , wherein each oligonucleotide sequence is RNA, the nucleotide sequence is shown in Table 1, and the target is mRNA. Among them, the sequence of lowercase letter...
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