Use of high efficiency siRNA for preparing drug capable of adjusting and controlling tumor cell cycle
A tumor cell, cycle technology, applied in the field of biomedicine, can solve problems such as loss of resistance
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0021] siRNA design and synthesis
[0022] Instruments OligoPilot II DNA / RNA Synthesizer (Pharmacia, Sweden), HP-1100 High Performance Liquid Chromatography (Hewlett-Packard, USA), DU 2640 UV Spectrophotometer (Backman, USA).
[0023] Materials and Reagents Synthesis column (6.3mL), molecular sieve (3A Sigma), 30HL carrier (lot number 256723), RNA Pac PA100 analysis column (Dionex, USA), 5 kinds of nucleotide monomers (A, U, C, G, dT), acetonitrile, dichloroacetic acid, tetrazole, N2 methylimidazole, except the manufacturers indicated, the rest are products of Pharmacia, Sweden.
[0024]Design of siRNA According to the sequence of the open reading frame (ORF) of the survivin gene, the small molecule interference RNA CPUsiRNA3 and 1 pair of negative control siRNA were designed, and their sequences were compared in the human EST database to determine that there were no other gene sequences identical to them. CPUsiRNA3 sense strand 5'-AGCAUUCGUCCGGUUGCGCtt-3', antisense strand 3...
Embodiment 2
[0029] Antitumor Pharmacodynamics Test
[0030] Materials and methods:
[0031] Lung cancer A549, gastric cancer BGC-823, liver cancer Bel7402, breast cancer MDA-MB-435, and cervical cancer Hela cell lines were preserved in our laboratory. The transfection reagent Lipofectamin TM2000 was purchased from Invitrogen, Taq DNA polymerase, dNTP, DNA Marker and protein Marker was purchased from Takara Corporation. Rabbit anti-human antibody of Survivin protein, HRP-goat anti-rabbit IgG antibody, and DAB chromogenic kit were purchased from Wuhan Boster Company.
[0032] Cell culture Lung cancer A549, gastric cancer BGC-823, liver cancer Bel7402, breast cancer MDA-MB-435, and cervical cancer Hela cell lines were cultured in RPMI1640 medium containing 10% fetal bovine serum in an incubator at 37°C and 5% CO2 .
[0033] MTT method will 2.5×10 6 / L cells were inoculated in a 96-well plate, 180 μl per well, 3 wells in parallel for each group, and a blank and negative drug control were ...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com