WO2018170762A1 - Vecteur d'expression d'arni du gène tnlg5a, procédé de construction associé et application correspondante - Google Patents
Vecteur d'expression d'arni du gène tnlg5a, procédé de construction associé et application correspondante Download PDFInfo
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- WO2018170762A1 WO2018170762A1 PCT/CN2017/077604 CN2017077604W WO2018170762A1 WO 2018170762 A1 WO2018170762 A1 WO 2018170762A1 CN 2017077604 W CN2017077604 W CN 2017077604W WO 2018170762 A1 WO2018170762 A1 WO 2018170762A1
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- sequence
- gene
- tnlg5a
- expression vector
- vector
- Prior art date
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Definitions
- a polynucleotide represented by SEQ ID NO: 1 or a vector comprising the polynucleotide represented by SEQ ID NO: 1 or an RNAi expression vector of the aforementioned TNLG5A gene is provided in TNLG5A Use in drugs for diseases with abnormal gene expression.
- RNAi expression vector of the TNLG5 A gene provided by the invention has the advantages of high transfection efficiency, low dosage, high and specific inhibition of TNLG5A gene expression in Jurkat cells, and can be used as a powerful tool for preparing and treating TNLG5A gene expression abnormality. Drugs related to the disease.
- FIG. 1 is a schematic diagram showing the results of real-time quantitative PCR detection of RNAi expression vector cells transfected with TNLG5A gene.
- Jurkat cells were purchased from the Cell Resource Center of the Shanghai Institute of Biological Sciences.
- the RNAi vector pLVX-shRNA1 was purchased from C1 ontech, RNeasy MiniKit was purchased from Qiagen, and the endotoxin-free plasmid extraction kit was purchased from Ome ga bio-tek.
- the complete medium described below is a cell culture medium supplemented with 10% fetal bovine serum.
- Synthetic shRNAs are designed based on the target nucleotide sequence.
- the design is as follows: B a m HI cleavage site +19 n t target nucleotide sequence + stem loop structure (TTCAAGAGA) + target sequence complementary sequence + RNAPolym polymerase transcriptional stop site (TTTTTT) + EcoR I cleavage site Six regions, the sequence of which is shown in SEQ ID NO: 1.
- the designed shRNA oligonucleotide chain was synthesized by Shanghai Shenggong Bioengineering Technology Service Co., Ltd.
- Jurkat cells were cultured, and 5,000,000 cells in good growth state were taken, and the cells were collected by centrifugation, and then resuspended in 500 ⁇ L of PBS, mixed with the RNAi expression vector of 20 ⁇ ⁇ TNLG5 ⁇ gene, and then added to an electric shock cup, and BTX ECM830 was applied.
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
L'invention concerne un vecteur d'expression d'ARNi du gène TNLG5A, un procédé de construction associé et une application correspondante. L'invention concerne : un polynucléotide isolé présentant une séquence telle que représentée dans la séquence SEQ ID NO:1 ; un vecteur contenant le polynucléotide tel que représenté par la séquence SEQ ID NO:1 ; un vecteur recombinant contenant une séquence telle que représentée dans la séquence SEQ ID NO:2 ; et un vecteur d'expression d'ARNi du gène TNLG5A et un procédé de préparation associé. Le procédé de préparation comprend : l'utilisation de pLVX-shARN1 en tant que vecteur de squelette et l'insertion dans un site recombinant d'une séquence oligonucléotidique de shARN (ARN court en épingle à cheveux) consistant en un site de coupe d'enzyme de Bam HI + une séquence nucléotidique cible + une séquence de structure tige-boucle + une séquence complémentaire à une séquence nucléotidique cible + une séquence de site de terminaison + un site de coupe d'enzyme EcoR I en liaison séquentielle.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2017/077604 WO2018170762A1 (fr) | 2017-03-22 | 2017-03-22 | Vecteur d'expression d'arni du gène tnlg5a, procédé de construction associé et application correspondante |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2017/077604 WO2018170762A1 (fr) | 2017-03-22 | 2017-03-22 | Vecteur d'expression d'arni du gène tnlg5a, procédé de construction associé et application correspondante |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2018170762A1 true WO2018170762A1 (fr) | 2018-09-27 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/CN2017/077604 WO2018170762A1 (fr) | 2017-03-22 | 2017-03-22 | Vecteur d'expression d'arni du gène tnlg5a, procédé de construction associé et application correspondante |
Country Status (1)
| Country | Link |
|---|---|
| WO (1) | WO2018170762A1 (fr) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101948859A (zh) * | 2010-08-17 | 2011-01-19 | 深圳市疾病预防控制中心 | 特异性抑制SET蛋白表达的shRNA表达载体及其构建方法和应用 |
-
2017
- 2017-03-22 WO PCT/CN2017/077604 patent/WO2018170762A1/fr active Application Filing
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101948859A (zh) * | 2010-08-17 | 2011-01-19 | 深圳市疾病预防控制中心 | 特异性抑制SET蛋白表达的shRNA表达载体及其构建方法和应用 |
Non-Patent Citations (1)
| Title |
|---|
| WANG, HUI ET AL.: "4-1BBL Small Interfering RNA Inhibits the Proliferation of HL-60B Cell in Vitro", JOURNAL OF BENGBU MEDICAL COLLEGE, vol. 2010, no. 8, 15 August 2010 (2010-08-15), pages 757 - 761 * |
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