Small interfering RNA capable of treating cancer
A chain structure, single-stranded technology, applied in DNA/RNA fragments, recombinant DNA technology, medical preparations containing active ingredients, etc., can solve the problem of low serological stability, achieve high application value, and improve reproductive inhibition rate , the effect of improving stability
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0047] Example 1: Design and synthesis of siRNA
[0048] Firstly, 300 siRNAs were designed according to the template of SEQ ID NO:11. For the above-mentioned 300 siRNAs obtained through design, further optimize. Finally, 4 siRNA sequences were selected, and the results are shown in Table 1. The oligonucleotide single strand of siRNA is chemically synthesized according to methods known in the art. The sequences of the synthesized oligonucleotides are shown in Table 1. During synthesis, two deoxythymine nucleotides dTdT are added to the 3' end of the single-stranded oligonucleotide. The siRNA is modified, and the modification is also obtained through numerous modification screens. Specifically, Sangon Bioengineering (Shanghai) Co., Ltd. was commissioned to synthesize the oligonucleotide sequences described in Table 1 below. The modification (OMe) means that the 2' hydroxyl group of the pentose group in the nucleotide residue to its left is replaced by a methoxy group. (F) ...
Embodiment 2
[0052] Example 2: Evaluation of the Effect of siRNA on Serum Stability
[0053] DD3-137, DD3-256, DD3-335, DD3-424, DD3-D, DD3-137(X), DD3-335(X) obtained in Preparation Example 1 were tested for their stability in serum environment. Specific steps are as follows.
[0054] Mix 10 μl of the above-mentioned modified and unmodified siRNA (20 mmol) with 50 μl of fetal bovine serum (Gibco fetal bovine serum (Gibco: 16000-044) and 40 μl of PBS, respectively, and incubate at 37°C for 0, 2, 4, 8, 24 , after 48 and 72 hours, obtain the processing sample. Processing sample sampling 10 μ l carries out 20% PAGE gel electrophoresis. The degradation rate is calculated by the ratio of the electrophoresis band light intensity of the above-mentioned treatment sample and the electrophoresis band light intensity of 0 hours, the result is as follows Shown in table 2.The degradation rate listed in table 2 is the degradation rate calculated by the ratio of the electrophoresis band light intensity ...
Embodiment 3
[0057] Example 3 siRNA verification of cell inhibitory effect
[0058]LNCaP clone FGC (human prostate cancer cell) product number: CL-0143, purchased from Wuhan Punuosheng Life Science and Technology Co., Ltd. Cell culture was carried out according to the instructions, and the cell density was 1X 105 cells / mL. Human normal prostate epithelial cells RWPE-1 (Catalog No.: CL-0200) were used as control cells, and the cell culture was carried out according to the instructions, and the cell density was 1X 105 / mL
[0059] 1X 10 per well 4 Inoculate a 96-well plate with 4 cells, infect siRNA, NC negative control, and set up a blank control group (control), and then continue to culture for about 24 hours, 48 hours, and 72 hours. Add 50 μl of 1 × MTT solution to each well and put it into the incubator. 4h to restore MTT. Aspirate the supernatant, add 150 μL DMSO to each well, and shake well with a plate shaker. Measure the optical density of each hole with a microplate reader at 4...
PUM
| Property | Measurement | Unit |
|---|---|---|
| Sensitivity | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More