WO2019033246A1 - Arn tud pour la co-invalidation génique de trois miarn et utilisation associée - Google Patents
Arn tud pour la co-invalidation génique de trois miarn et utilisation associée Download PDFInfo
- Publication number
- WO2019033246A1 WO2019033246A1 PCT/CN2017/097429 CN2017097429W WO2019033246A1 WO 2019033246 A1 WO2019033246 A1 WO 2019033246A1 CN 2017097429 W CN2017097429 W CN 2017097429W WO 2019033246 A1 WO2019033246 A1 WO 2019033246A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- mir
- tud
- vector
- pglv3
- preparation
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/66—General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
Definitions
- miR -185 is a 22 nt miRNA, located in human chromosome 22ql l.21, plays an important role as a tumor suppressor gene in the development and invasion of tumors such as colon cancer, gastric cancer, esophageal cancer, lung cancer, liver cancer, etc.
- Tough Decoy RNA is a novel miRNA-inhibiting miRNA that inhibits miRNA by introducing double-stranded RNA to target miRNAs. Because the inserted RNA is double-stranded and has a secondary structure of stem-loops, it is resistant to intracellular nuclease degradation and inhibits miRNAs in a long-term, stable, and efficient manner.
- the RNA is constructed on a lentiviral vector to obtain a recombinant vector containing the Tud RNA; and the recombinant vector containing the Tud RNA is applied to 16HBE cells to inhibit the expression of miR-152, miR-185 and miR-424;
- the 5' end of the sense strand template is added with GATCC, which is complementary to the sticky end formed by BamHI digestion; the AATTC is added to the 5' end of the antisense strand template, which is complementary to the sticky end formed by EcoRI digestion.
- the cDNA of each of the two cells was used as a template.
- the expression levels of miR-152, miR-185 and miR-424 were detected by real-time PCR, and the experiment was repeated three times. Three parallel samples were set per well, and snord 44 was used as an internal reference. The results are shown in Figure 2. It can be seen that the expression level of miR-152 in TuD-152-185-424 cells is 58 ⁇ 3 ⁇ 4 lower than that in 16HBE cells, and the expression level of miR-185 is 49% lower than that in 16HBE cells, miR-424. The expression level was 68% lower than that of 16HBE cells. The difference was statistically significant ( ⁇ 0.01), indicating that the TuD-152-185-424 cell line was successfully constructed.
Landscapes
- Genetics & Genomics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
L'invention concerne un ARN Tud pour l'inactivation génique des miARN-29a, miARN-152, et miARN-424. L'invention concerne également un procédé de connexion d'une séquence nucléotidique codant pour l'ARN TuD à un vecteur navette lentiviral pour préparer un vecteur recombinant et son utilisation pharmaceutique.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/CN2017/097429 WO2019033246A1 (fr) | 2017-08-14 | 2017-08-14 | Arn tud pour la co-invalidation génique de trois miarn et utilisation associée |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/CN2017/097429 WO2019033246A1 (fr) | 2017-08-14 | 2017-08-14 | Arn tud pour la co-invalidation génique de trois miarn et utilisation associée |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2019033246A1 true WO2019033246A1 (fr) | 2019-02-21 |
Family
ID=65361632
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2017/097429 WO2019033246A1 (fr) | 2017-08-14 | 2017-08-14 | Arn tud pour la co-invalidation génique de trois miarn et utilisation associée |
Country Status (1)
Country | Link |
---|---|
WO (1) | WO2019033246A1 (fr) |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101368213A (zh) * | 2008-10-13 | 2009-02-18 | 南京大学 | 血清微小核糖核酸试剂盒及其在乙肝早期诊断中的应用 |
WO2010138263A2 (fr) * | 2009-05-28 | 2010-12-02 | University Of Massachusetts | Nouveaux virus adéno-associés (aav) et leurs utilisations |
CN102218144A (zh) * | 2010-04-13 | 2011-10-19 | 江苏命码生物科技有限公司 | 一种调节生物体内微小核糖核酸含量的方法及其用途 |
CN103080334A (zh) * | 2010-06-04 | 2013-05-01 | 复旦大学 | 用于诊断早期结直肠癌及高级腺瘤的微rna生物标记及方法 |
CN104531700A (zh) * | 2014-11-11 | 2015-04-22 | 西北工业大学 | 抑制小鼠MACF1基因表达的shRNA序列及其应用 |
-
2017
- 2017-08-14 WO PCT/CN2017/097429 patent/WO2019033246A1/fr active Application Filing
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101368213A (zh) * | 2008-10-13 | 2009-02-18 | 南京大学 | 血清微小核糖核酸试剂盒及其在乙肝早期诊断中的应用 |
WO2010138263A2 (fr) * | 2009-05-28 | 2010-12-02 | University Of Massachusetts | Nouveaux virus adéno-associés (aav) et leurs utilisations |
CN102218144A (zh) * | 2010-04-13 | 2011-10-19 | 江苏命码生物科技有限公司 | 一种调节生物体内微小核糖核酸含量的方法及其用途 |
CN103080334A (zh) * | 2010-06-04 | 2013-05-01 | 复旦大学 | 用于诊断早期结直肠癌及高级腺瘤的微rna生物标记及方法 |
CN104531700A (zh) * | 2014-11-11 | 2015-04-22 | 西北工业大学 | 抑制小鼠MACF1基因表达的shRNA序列及其应用 |
Non-Patent Citations (1)
Title |
---|
ZHU LINWENSI, FANG JINGYUAN: "The Structure and Clinical Roles of MicroRNA in Colorectal Cancer", GASTROENTEROLOGY RESEARCH AND PRACTICE, vol. 2016, 31 December 2016 (2016-12-31), pages 1 - 6, XP055540703, ISSN: 1687-630X * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP2925866B1 (fr) | Arn circulaire destiné à l'inhibition de micro-arn | |
CA3136735A1 (fr) | Procedes et compositions pour editer des arn | |
US20170004254A1 (en) | MULTI-TARGETING SHORT INTERFERING RNAs | |
CN107365785B (zh) | 一种调控细胞内NF-κB活性的基因表达载体及其调控方法和应用 | |
WO2019196887A1 (fr) | Nouveau petit arn activateur | |
JP2015212310A (ja) | ランダムRNAiライブラリ、その生成方法、及びそれを使用したスクリーニング方法 | |
JP2012528588A (ja) | 単一またはマルチ標的遺伝子を抑制するマルチ−シストロンshRNA発現カセット | |
JP6137484B2 (ja) | 遺伝子発現抑制用二本鎖核酸分子 | |
WO2012142480A1 (fr) | Oligonucléotides d'arn chimériques et leurs utilisations | |
WO2019033246A1 (fr) | Arn tud pour la co-invalidation génique de trois miarn et utilisation associée | |
WO2018165929A1 (fr) | Vecteur d'expression inhibant le micro-arn double brin, son procédé de mise au point et son application | |
JP2022531580A (ja) | ベクターでの使用のためのマルチプレックス化shRNA | |
WO2019036871A1 (fr) | Arn tud pour l'inactivation spécifique des expressions de mir-148a, mir-185 et mi-424 d'êtres humains, et applications associées | |
WO2017214952A1 (fr) | Construction et application d'un vecteur lentiviral pour l'inhibition spécifique de l'expression de l'arnmi-185 humain | |
WO2019000147A1 (fr) | Arn tud inhibant efficacement l'expression des mir-148a, mir-152 et mir-185 humains et son utilisation | |
WO2019033248A1 (fr) | Arn tud inhibant l'expression des mir-148a, mir-152 et mir-424 humains et son utilisation | |
WO2017219166A1 (fr) | Vecteur lentiviral pour l'inhibition simultanée de l'expression de deux miarn, et application associée | |
WO2018170754A1 (fr) | Adénovirus ad-29a-152-424-tud recombinant, et construction et application de celui-ci | |
WO2018170620A1 (fr) | Vecteur d'expression de deux micro-arn mis sous silence | |
WO2018170758A1 (fr) | Adénovirus ad-140-148a-152-tud recombinant, et construction et application de celui-ci | |
WO2018170757A1 (fr) | Adénovirus ad-29a-185-424-tud recombinant, et construction et application associées | |
WO2018170759A1 (fr) | Adénovirus recombinant ad-140-148a-185-tud, construction et application de celui-ci | |
CN110923234A (zh) | 一种抑制人GLI2基因表达的shRNA构建及其应用 | |
WO2018170751A1 (fr) | Arn tud destiné à antagoniser l'expression d'un micro-arn, et son application | |
WO2018170653A1 (fr) | Arn tud destiné à diminuer les niveaux d'expression de miarn-29a, mir-140 et mir-424, et son application |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 17921814 Country of ref document: EP Kind code of ref document: A1 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 17921814 Country of ref document: EP Kind code of ref document: A1 |