Method for screening siRNA on basis of bioinformatics
A bioinformatics and gene technology, applied in the field of small nucleic acid drugs, to achieve the effect of avoiding spread, avoiding toxic and side effects, and rapid response
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Embodiment 1
[0087] The cRNA sequence (GeneBank accession number: KC853766.1) of a H7N9 influenza A virus genome HA protein was obtained from NCBI (National Center for Biotechnology Information, website: www.ncbi.nlm.nih.gov), with a length of 1689bp.
[0088] Using the cRNA sequence as a template, use the siDirect online design tool to design the first candidate siRNA sequence. Specifically, the cRNA sequence was input into the siDirect online design tool (URL: http: / / sidirect2.rnai.jp / ), and 67 first candidate siRNA sequences were obtained.
[0089] The obtained first candidate siRNA sequences were sorted according to the thermodynamic stability of the double helix formed by the seed region and the target sequence (seed duplex stability), and the highest stability (T m value ≤ 10°C), the first candidate siRNA sequence has 4 corresponding target site sequences:
[0090] Sequence 1: 5'-ACCAAAGTAAACACATTAACTGA-3'
[0091] Sequence 2: 5'-TAGAATTTTCAGCCGATTTAATT-3'
[0092] Sequence 3: 5'-...
Embodiment 2
[0100] The cRNA sequence (GeneBank accession number: KC853766.1) of a H7N9 influenza A virus genome HA protein was obtained from NCBI (National Center for Biotechnology Information, website: www.ncbi.nlm.nih.gov / ), with a length of 1689bp .
[0101] Using the cRNA sequence as a template, the first candidate siRNA sequence was designed using siDirect, Dharmacon, and GenScript online design tools, respectively. Specifically, the cRNA sequence was input into (1) siDirect online design tool (URL: http: / / sidirect2.rnai.jp / ), (2) Dharmacon online design tool (URL: http: / / www.thermoscientificbio.com / design-center / ?redirect=true) and (3) GenScript online design tool (URL: https: / / www.genscript.com / ssl-bin / app / rnai), get the first candidate siRNA sequence and The ranking results are compared and those first candidate siRNA sequences with higher comprehensive scores are selected.
[0102] Eliminate the first candidate siRNA sequence that can produce off-target effects on human genes ...
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