SiRNA inhibiting PPFIA1 gene expression, and its application
A gene expression and gene technology, applied in the field of siRNA inhibiting PPFIA1 gene expression
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[0030] (1) Design and synthesis of PPFIA1siRNA:
[0031] According to the known sequence of PPFIA1 in the GenBank gene bank (Gene ID: 8500, NM_003626.2, NM_177423.1), according to the siRNA design principle, combined with the online design tool siCatch provided by Ambion TM siRNA Design is designed for the siRNA target position in the coding region;
[0032] The sequence of PPFIA1 siRNA is:
[0033] Sense strand: 5'-CCACAAAGCUCUGGAUGAA-3',
[0034] Antisense strand: 5'-UUCAUCCAGAGCUUUGUGG-3';
[0035] There is dTdT overhanging at the 3' end of the PFIA1 siRNA sequence, which was synthesized by Guangzhou Ruibo Biotechnology Co., Ltd.
[0036] The random sequence RNA duplex (scramble, SCR) was used as a random control, and the random sequence RNA duplex was purchased from Shanghai Gemma Pharmaceutical Technology Co., Ltd., and its catalog number is: A06001.
[0037] (2) Culture of leukemia K562 cells
[0038] Leukemic K562 cells were inoculated in 1640 medium containing 10...
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