Interference of transgene mouse model by slow viral vectors mediation IKK alpha gene RNA and use thereof
A technology of transgenic mice and α gene, which is applied in the field of medicine and medical treatment, and can solve problems such as irreplaceable
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[0092] Example: Preparation method of lentiviral vector-mediated IKKα gene RNA interference transgenic mice
[0093] 1. Synthesis of two oligonucleotide chains.
[0094] According to the online RNAi sequence design software of Invitrogen Company of the United States, search for the sequence of the knock-down IKKα gene, and synthesize two oligonucleotide chains, the sequences of which are as follows:
[0095] 5′-CACCGCAGGCTCTTTCAGGGACACGAATGTCCCTGAAAGAGCCTGC-3′
[0096] 5′-AAAAGCAGGCTCTTTCAGGGACATTCGTGTCCCTGAAAGAGCCTGC-3′
[0097] An annealing reaction was then carried out, and the annealed product was named IKKαds Oligo, and the structure after annealing was as follows:
[0098]
[0099] 2. Preparation of expression vector (pU6-IKKα-knockdown expression vector).
[0100]The above IKKαds Oligo was ligated with 2 μl pENT / U6vector (0.5ng / μl) to transform TOP 10 competent bacteria, ligated with 2 μl pENT / U6vector (0.5ng / μl) to transform TOP10 competent bacteria, and then the...
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