WO2017214943A1 - Vecteur d'expression de lentivirus pour favoriser l'expression du gène tim-3 et son application - Google Patents
Vecteur d'expression de lentivirus pour favoriser l'expression du gène tim-3 et son application Download PDFInfo
- Publication number
- WO2017214943A1 WO2017214943A1 PCT/CN2016/086067 CN2016086067W WO2017214943A1 WO 2017214943 A1 WO2017214943 A1 WO 2017214943A1 CN 2016086067 W CN2016086067 W CN 2016086067W WO 2017214943 A1 WO2017214943 A1 WO 2017214943A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- tim
- gene
- sequence
- vector
- expression
- Prior art date
Links
- 230000014509 gene expression Effects 0.000 title claims abstract description 44
- 239000013604 expression vector Substances 0.000 title claims abstract description 33
- 102100034458 Hepatitis A virus cellular receptor 2 Human genes 0.000 title claims abstract description 11
- 230000001737 promoting effect Effects 0.000 title claims abstract description 4
- 101001068133 Homo sapiens Hepatitis A virus cellular receptor 2 Proteins 0.000 title claims 2
- 101710083479 Hepatitis A virus cellular receptor 2 homolog Proteins 0.000 claims abstract description 80
- 239000002299 complementary DNA Substances 0.000 claims abstract description 29
- 102000004190 Enzymes Human genes 0.000 claims abstract description 15
- 108090000790 Enzymes Proteins 0.000 claims abstract description 15
- 229940126547 T-cell immunoglobulin mucin-3 Drugs 0.000 claims abstract description 15
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 11
- 108091026890 Coding region Proteins 0.000 claims abstract description 10
- 238000010367 cloning Methods 0.000 claims abstract description 10
- 239000013598 vector Substances 0.000 claims description 23
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 claims description 13
- 229960000723 ampicillin Drugs 0.000 claims description 13
- 238000011144 upstream manufacturing Methods 0.000 claims description 13
- 239000013612 plasmid Substances 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 11
- 238000012163 sequencing technique Methods 0.000 claims description 9
- 241000588724 Escherichia coli Species 0.000 claims description 8
- 230000001580 bacterial effect Effects 0.000 claims description 7
- 238000010276 construction Methods 0.000 claims description 7
- 102000012410 DNA Ligases Human genes 0.000 claims description 5
- 108010061982 DNA Ligases Proteins 0.000 claims description 5
- 238000001962 electrophoresis Methods 0.000 claims description 5
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 4
- 238000012408 PCR amplification Methods 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 238000013461 design Methods 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 4
- 108091008146 restriction endonucleases Proteins 0.000 claims description 4
- 230000002159 abnormal effect Effects 0.000 claims description 3
- 201000010099 disease Diseases 0.000 claims description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 3
- 229940079593 drug Drugs 0.000 claims description 3
- 239000012634 fragment Substances 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 230000003321 amplification Effects 0.000 claims description 2
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 2
- 108020004414 DNA Proteins 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 29
- 239000002609 medium Substances 0.000 description 14
- 241000700605 Viruses Species 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 239000013642 negative control Substances 0.000 description 8
- 239000013641 positive control Substances 0.000 description 8
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 8
- 206010028980 Neoplasm Diseases 0.000 description 6
- 238000003776 cleavage reaction Methods 0.000 description 6
- 230000007017 scission Effects 0.000 description 6
- 210000001744 T-lymphocyte Anatomy 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 4
- 229950010131 puromycin Drugs 0.000 description 4
- 101150090724 3 gene Proteins 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 238000004806 packaging method and process Methods 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 230000002459 sustained effect Effects 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 238000001890 transfection Methods 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 2
- 229930182555 Penicillin Natural products 0.000 description 2
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 235000010633 broth Nutrition 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 239000000539 dimer Substances 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 2
- 229940049954 penicillin Drugs 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000012827 research and development Methods 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 210000004981 tumor-associated macrophage Anatomy 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 102000015212 Fas Ligand Protein Human genes 0.000 description 1
- 108010039471 Fas Ligand Protein Proteins 0.000 description 1
- 101150046249 Havcr2 gene Proteins 0.000 description 1
- 229920000209 Hexadimethrine bromide Polymers 0.000 description 1
- 102000008949 Histocompatibility Antigens Class I Human genes 0.000 description 1
- 108010088652 Histocompatibility Antigens Class I Proteins 0.000 description 1
- 238000011530 RNeasy Mini Kit Methods 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- 230000006044 T cell activation Effects 0.000 description 1
- 230000033540 T cell apoptotic process Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 210000000612 antigen-presenting cell Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000020411 cell activation Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 230000003828 downregulation Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 210000003289 regulatory T cell Anatomy 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
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
Definitions
- the present invention belongs to the field of genetic engineering technology, and in particular relates to a lentiviral expression vector which promotes expression of a TIM-3 gene and an application thereof.
- Tumors can be used to strengthen the "army” by various methods: 1) Tumor cells have low expression or lack of expression of specific or related antigens, and down-regulate MHC-I molecules by gene defect or transcription down-regulation, and antigen-presenting cells cannot give T
- B7 molecule such as B7-l, B7-2 or up-regulation of B7-H1, B7-H2, etc., such that T cell activation is deficient in the second signal or induces T cell inactivation, Apoptosis
- Tumor cells can induce autologous apoptosis or induce T cell apoptosis by low expression of Fas and defective Fas signaling molecules or high expression of FasL
- 4) release of tumor-derived factors and induce regulatory T cells (Treg) , Regulatory DCs (DCreg), tumor-associated macrophages (TAM), long-term stimulation of a large number of tumor antigens and immune molecules, leading to T cell depletion.
- DCreg regulatory DCs
- TAM tumor-associated macrophages
- Negative co-stimulatory molecules PD-1 and Tim3 are important marker molecules for early and late viral responsive CD8+ T cell depletion, respectively. Therefore, the study of TIM-3 is expected to enhance the ability of T cells to kill tumor cells and rescue depleted T cells. However, the lack of lentiviral expression vectors that specifically promote the high expression of TIM-3 gene in the prior art makes the related studies not well. exhibition.
- the TIM-3 gene cDNA sequence of the I cleavage site was inserted into the multiple cloning site of the pLVX-IRES-Puro expression vector to construct a lentiviral expression vector which specifically promotes high expression of the TIM-3 gene, thereby completing the present invention.
- the present invention provides a lentiviral expression vector which specifically promotes high expression of a TIM-3 gene, including a basic sequence of a pLVX-IRE S-puro expression vector, a resistance gene sequence, a multiple cloning site sequence, a promoter sequence, and TI M-3 gene cDNA sequence; the multiple cloning site includes Spe I cleavage site and Not
- the TIM-3 gene cDNA sequence includes a Spe l cleavage site, a TIM-3 gene coding sequence and a Not I cleavage site, and the TIM-3 gene cDNA sequence is inserted into the poly Cloning of the site sequence.
- the lentiviral expression vector constructed by inserting the cDNA sequence of the TIM-3 gene into the pLVX-IRES-Puro expression vector has the advantages of high transfection efficiency, low dosage, sustainable, high efficiency and stability.
- the advantages of increasing the expression of TIM-3 gene can be used as a powerful tool for the preparation and treatment of TIM-3 gene expression in the treatment of diseases such as Alzheimer's disease.
- the TIM-3 gene coding sequence is obtained by PCR amplification
- the PCR primer comprises an upstream primer and a downstream primer
- the sequence of the upstream primer is: 5'-GACTAGTATGTTTTCACATCTTCCCTTTG -3', ie SEQ ID NO: 1
- the sequence of the downstream primer is: 5, - GGCGGCCGCTGTCGCTTTGCAATGCCATAG -3', ie SEQ ID NO: 2.
- the TIM-3 gene coding sequence can be amplified by PCR, and can be successfully inserted into the pLVX-IRES-Puro expression vector to continuously express the TIM-3 gene, which reduces the cost of sequence synthesis and lowers the cost.
- the present invention also provides a method for constructing a lentiviral expression vector which specifically promotes high expression of the TIM-3 gene, comprising the following steps:
- A) TIM-3 gene primer design According to the TIM-3 gene coding sequence, using Oligo 7 analysis, select 5'-GACTAGTATGTTTTCACATCTTCCCTTTG-3', ie SEQ ID NO: 1 as the upstream bow, select 5 '- GGCGGCCGCTGTCGCTTTGCAATGCCATAG -3, ie, SEQ ID NO: 2 as a downstream primer, then synthesizing the upstream primer and the downstream primer; the upstream primer and the downstream primer are free of primer dimer, and the annealing temperature difference is small;
- the successful bacterial liquid was sequenced and identified; the correct Escherichia coli was identified by liquid LB medium, and the pGM-T vector carrying the TIM-3 gene cDNA sequence was extracted, and the restriction endonuclease Spe I enzyme and Not I enzyme double digestion, electrophoresis, gelatinization to recover a fragment of about 1000 bp, this fragment is the TIM-3 gene cDNA sequence;
- the present invention utilizes genetic engineering technology to construct a lentiviral expression vector which specifically promotes high expression of TIM-3 gene. After successful identification, it is packaged into a virus and introduced into Jurkat cells. After screening cells with puromycin, the real-time fluorescence is quantified. The PCR and Western Blot techniques verified the change of TIM-3 gene expression from mRNA and protein levels, respectively. The experimental results confirmed that the TIM-3 gene cDNA sequence provided by the present invention was successfully inserted into the pLVX-I RES-Puro expression vector, which can be specific and sustained. Highly and stably promote high expression of TIM-3 gene.
- the present invention also provides a use of a lentiviral expression vector which specifically promotes high expression of a TIM-3 gene for the preparation of a medicament for treating a disease associated with abnormal expression of a TIM-3 gene.
- the lentiviral expression vector which specifically promotes the high expression of the TIM-3 gene provided by the invention has the advantages of high transfection efficiency, low dosage, and can promote the high expression of the TIM-3 gene in a specific, sustained, efficient and stable manner.
- the present invention also provides specific promotion of TIM-
- the construction method of the lentiviral expression vector with high expression of 3 gene has good operation effect, reduces the cost of sequence synthesis, and has low cost.
- 1 is a plasmid map of the pLVX-IRES-Puro expression vector.
- FIG. 2 is a schematic diagram showing the results of fluorescent quantitative PCR detection after puromycin screening of cells.
- Jurkat cells were purchased from the Cell Resource Center of the Shanghai Institute of Biological Sciences. 293FT cells were purchased from Thermo Fisher, Premix PrimeSTAR HS enzyme, lentiviral expression vector pLVX-IRES-Puro, virus packaging auxiliary kit, and Lenti-X GoStix kit. Purchased from Takara, RNeasy Mini
- Kit was purchased from QIAGEN, pGM-T was purchased from Tiangen, and Endo-Free Plasmid Mini Kit II was purchased from Omega bio-tek.
- TIM-3 gene coding sequence GenBank NMJ 2782.4
- Example 2 Construction of a lentiviral vector that specifically promotes high expression of the TIM-3 gene
- the coding sequence of the TIM-3 gene was sequenced with Premix PrimeSTAR HS enzyme. Amplification, electrophoresis recovery and then adding A tail reaction, using T4
- the DNA ligase was ligated to the pGM-T vector to obtain the ligation product (TIM-3-T vector), and the ligation product was transformed into competent E. coli DH5CC and uniformly coated on an ampicillin-containing LB medium plate at 37 Cultured at °C for 12 h, the same control group was set to negative control group 1 (the competent cells were uniformly coated on the ampicillin-free plate), and the negative control group 2 (the competent cells were uniformly coated with 100 g/ml ampicillin).
- positive control group 1 the connection product of the double enzyme-cut empty vector was uniformly coated on the plate containing 100 g/ml ampicillin
- positive control group 2 the empty carrier was uniformly coated in 100%
- the bacteria with the correct sequencing result were cultured in liquid LB medium for 14 h, and then the recombinant T vector containing the TIM-3 gene sequence was extracted, and the pLVX-IRES-Puro vector was firstly treated with Spe I enzyme and The Not I enzyme was double-digested, electrophoresed and recovered, and the product was recovered by T4 DNA ligase, and then transformed into competent E. coli DH50C, uniformly coated on an ampicillin-containing LB medium plate, and cultured at 37 ° C.
- the same negative control group 1 was set (the competent cells were uniformly coated on the ampicillin-free plate), and the negative control group 2 (the competent cells were uniformly coated on the plate containing 100 g/ml ampicillin) ), positive control group 1 (the ligation product of the double enzyme-cut empty vector was uniformly coated on a plate containing 100 g/ml ampicillin), positive control group 2 (the empty carrier was uniformly coated in 100 g/mL ampicillin) Penicillin on the plate).
- the experimental group grew a single colony, and the negative control group 1 grew colonies; the negative control group 2, the positive control group 1, and the positive control group 2 did not grow colonies.
- 293FT cells were cultured, and cells grown in good condition were inoculated into six wells, 1,000,000 cells per well, and the extracted recombinant plasmid pLVX-TIM-3 2 ⁇ ⁇ was transfected into 293FT using a lentiviral packaging auxiliary kit. After 48 h, the virus-containing supernatant medium was collected, and the virus solution was filtered through a sieve of 0.45 ⁇ m for infecting Jur kat cells, and the Lenti-X GoStix kit was used to detect the virus titer of 5000000 to 50000000 IFU.
- Jurkat cells were inoculated into 6-well plates at 1000000 cells per well. After 12 hours, the cell density was about 50 ⁇ 3 ⁇ 4, and the virus solution was taken separately. The virus was diluted 10 times with DMEM complete medium, and polybrene was added. ) The final concentration is 8 g/mL. The medium in the 6-well plate was added to the virus-containing DMEM complete medium (containing 10% fetal bovine serum). After 24 hours, the virus-containing DMEM complete medium was discarded, and the fresh DMEM complete medium was replaced. After 24 hours, 0.5 was used. The cells were screened at a g/ml concentration of puromycin. After 10 days of screening, the medium was changed once every 3 days, and the concentration of puromycin was continuously increased to 1.00 g/ml.
- Example 5 Fluorescence quantitative PCR was used to detect the expression level of TIM-3 gene.
- the primer design software Oligo 7.0 was used to design the bow.
- Kit reverse transcribes mRNA into cDNA, reverse transcription conditions: 37 ° C, 15 min; 85 ° C, 5 s; 4 ° C, ⁇ . After the end of reverse transcription, add 9 (L of RNase Free dH 2 0 diluted cDNA, and store at -20 ° C for later detection. Take the cDNA of each group of cells.
- ⁇ was used as a template, and GAPDH was used as an internal reference.
- Real-time quantitative PCR was used to detect the relative expression of ⁇ -3, and the reaction conditions were set: 95. C 30s, 1 cycle, 54°C 30s 40 cycles, 95. C 5s, 60.
- TIM-3 gene is more than 60-fold higher than that of Jurkat cells, whether it has just been screened or has been cultured for 20 generations of pLVX-TIM-3 cells, whereas pLVX empty vector cells
- the expression level of TIM-3 gene was almost unchanged compared with Jurkat cells, indicating that the TIM-3 gene cDNA sequence provided by the present invention was successfully inserted into the pLVX-IRES-Puro expression vector, and the TIM- can be specifically, continuously, efficiently and stably promoted. 3 gene high expression.
- the lentiviral expression vector which specifically promotes the high expression of the TIM-3 gene provided by the invention has the advantages of high transfection efficiency, low dosage, and can promote the high expression of the TIM-3 gene in a specific, sustained, efficient and stable manner.
- the present invention also provides specific promotion of TIM-
- the construction method of the lentiviral expression vector with high expression of 3 gene has good operation effect, reduces the cost of sequence synthesis, and has low cost.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
L'invention porte sur un vecteur d'expression de lentivirus pour favoriser une expression plus élevée des gènes TIM-3, comprenant une séquence fondamentale, une séquence de gène de résistance, une séquence de site de clonage multiple, une séquence de promoteur et une séquence d'ADNc du gène TIM-3 d'un vecteur d'expression pLVX-IRES-puro. Un site de clonage multiple comprend un site de coupure de l'enzyme Spe I et un site de coupure de l'enzyme Not I; la séquence d'ADNc du gène TIM-3 comprend le site de coupure de l'enzyme Spe I, une séquence de codage TIM-3 et le site de coupure de l'enzyme TIM-3. La séquence d'ADNc du gène TIM -3 est insérée vers l'avant dans la séquence de site de clonage multiple.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2016/086067 WO2017214943A1 (fr) | 2016-06-16 | 2016-06-16 | Vecteur d'expression de lentivirus pour favoriser l'expression du gène tim-3 et son application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2016/086067 WO2017214943A1 (fr) | 2016-06-16 | 2016-06-16 | Vecteur d'expression de lentivirus pour favoriser l'expression du gène tim-3 et son application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2017214943A1 true WO2017214943A1 (fr) | 2017-12-21 |
Family
ID=60663821
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/CN2016/086067 WO2017214943A1 (fr) | 2016-06-16 | 2016-06-16 | Vecteur d'expression de lentivirus pour favoriser l'expression du gène tim-3 et son application |
Country Status (1)
| Country | Link |
|---|---|
| WO (1) | WO2017214943A1 (fr) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003063792A2 (fr) * | 2002-01-30 | 2003-08-07 | The Brigham And Women's Hospital, Inc. | Compositions et methodes associees a tim-3, molecule de surface cellulaire specifique a th1 |
| CN103079644A (zh) * | 2010-06-11 | 2013-05-01 | 协和发酵麒麟株式会社 | 抗tim-3抗体 |
-
2016
- 2016-06-16 WO PCT/CN2016/086067 patent/WO2017214943A1/fr active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003063792A2 (fr) * | 2002-01-30 | 2003-08-07 | The Brigham And Women's Hospital, Inc. | Compositions et methodes associees a tim-3, molecule de surface cellulaire specifique a th1 |
| CN103079644A (zh) * | 2010-06-11 | 2013-05-01 | 协和发酵麒麟株式会社 | 抗tim-3抗体 |
Non-Patent Citations (2)
| Title |
|---|
| SHE, JING: "The Effect of Tim-3 on TLR/LPS Signalling Pathway in Murine Macrophages", CHINESE MASTER'S THESES FULL-TEXT DATABASE MEDICINE AND HEALTH SCIENCE, 15 April 2014 (2014-04-15), pages 12 - 19 * |
| ZHANG, SHENGTAO ET AL.: "Expression of Human T- cell Immunoglobulin Mucin 3(TIM-3) on Eukaryotic Cells and Establishment of Stable Transfectant Cell Line", CHINESE JOURNAL OF CELLULAR AND MOLECULAR IMMUNOLOGY, vol. 21, no. 6, 31 December 2005 (2005-12-31), pages 707 - 709 and 713 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2019176869A5 (fr) | ||
| JP2022137112A (ja) | 内因性ｔ細胞受容体の標的置換 | |
| EP3765094A1 (fr) | Compositions de régulation génique et procédés pour améliorer l'immunothérapie | |
| AU2019236204A1 (en) | Gene-regulating compositions and methods for improved immunotherapy | |
| AU2019234926A1 (en) | Gene-regulating compositions and methods for improved immunotherapy | |
| CN110799644A (zh) | Cblb核酸内切酶变体、组合物及使用方法 | |
| CA3149897A1 (fr) | Methodes et compositions d'integration genomique | |
| WO2017101244A1 (fr) | Procédé de préparation et d'utilisation de vecteurs d'expression lentiviraux, et procédé de préparation de lentivirus recombinants | |
| WO2018006880A1 (fr) | Co-expression d'un récepteur de point de contrôle immunitaire recombinant et d'un inhibiteur du point de contrôle immunitaire et application | |
| JP2023516300A (ja) | 腫瘍浸潤リンパ球の活性化及び増殖のための方法 | |
| JP2023502780A (ja) | 腫瘍浸潤リンパ球の活性化及び増殖方法 | |
| IL297881A (en) | Selection by knock-in of essential genes | |
| CN103352052B (zh) | 一种多顺反子双标表达慢病毒载体构建及应用 | |
| WO2017214943A1 (fr) | Vecteur d'expression de lentivirus pour favoriser l'expression du gène tim-3 et son application | |
| US20220042048A1 (en) | Nonviral generation of genome edited chimeric antigen receptor t cells | |
| WO2017214940A1 (fr) | Vecteur d'expression lentiviral pour favoriser spécifiquement l'expression élevée du gène cplx2, et ses applications | |
| WO2017101243A1 (fr) | Procédé de préparation et d'utilisation de vecteurs d'expression lentiviraux, et procédé de préparation de lentivirus recombinants | |
| IL308445A (en) | Methods and compositions for genomic integration | |
| WO2017214829A1 (fr) | Vecteur d'expression de lentivirus pour favoriser spécifiquement l'expression élevée du gène pd-l1, et ses applications | |
| WO2017214828A1 (fr) | Vecteur d'expression lentiviral pour favoriser spécifiquement l'expression élevée du gène prkcz, et ses applications | |
| Miri et al. | Construction of bacterial ghosts for transfer and expression of a chimeric hepatitis C virus gene in macrophages | |
| WO2017214944A1 (fr) | Vecteur lentiviral pour favoriser l'expression supérieure de gènes tigit et ses applications | |
| WO2017214834A1 (fr) | Vecteur d'expression lentiviral pour favoriser spécifiquement l'expression élevée du gène ctla-4, et ses applications | |
| WO2017214830A1 (fr) | Vecteur lentiviral pour favoriser spécifiquement l'expression élevée du gène pd-1, et ses applications | |
| WO2017214942A1 (fr) | Vecteur d'expression lentiviral pour améliorer l'expression du gène tctp, et ses applications |
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: 16905058 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: 16905058 Country of ref document: EP Kind code of ref document: A1 |