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WO1998033903A1 - Promoteurs specifiques de tissu - Google Patents

Promoteurs specifiques de tissu Download PDF

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Publication number
WO1998033903A1
WO1998033903A1 PCT/AU1998/000057 AU9800057W WO9833903A1 WO 1998033903 A1 WO1998033903 A1 WO 1998033903A1 AU 9800057 W AU9800057 W AU 9800057W WO 9833903 A1 WO9833903 A1 WO 9833903A1
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Prior art keywords
promoter
enhancer
coding sequence
specific gene
genetic
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PCT/AU1998/000057
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English (en)
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Peter Laurence Molloy
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Commonwealth Scientific And Industrial Research Organisation
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Priority to CA002279552A priority Critical patent/CA2279552A1/fr
Priority to EP98901256A priority patent/EP1009820A1/fr
Priority to AU57421/98A priority patent/AU5742198A/en
Publication of WO1998033903A1 publication Critical patent/WO1998033903A1/fr

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    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6445—Kallikreins (3.4.21.34; 3.4.21.35)
    • 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
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • 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
    • C12N2830/00—Vector systems having a special element relevant for transcription
    • C12N2830/008—Vector systems having a special element relevant for transcription cell type or tissue specific enhancer/promoter combination
    • 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
    • C12N2830/00—Vector systems having a special element relevant for transcription
    • C12N2830/80—Vector systems having a special element relevant for transcription from vertebrates
    • C12N2830/85—Vector systems having a special element relevant for transcription from vertebrates mammalian

Definitions

  • the present invention is concerned with combinations of prostate gene promoters with a general enhancer, the SV40 enhancer, giving rise to essentially androgen-independent promoters active in prostate cells which are androgen-independent in their growth and survival.
  • PSBP rat prostatic steroid binding proteins
  • Pb probasin
  • prostatic acid phosphatase genes rat prostatic steroid binding proteins
  • the PSA and probasin regulatory regions are the two most studied among prostate-expressed genes. It has been established that a 430 bp region upstream of the rat probasin gene is able to confer prostate specificity of expression on a reporter gene (Greenberg, Demayo et al. 1994; Matusik; WO9403594); when used to target expression of the SV40 T-antigen, prostate tumours develop specifically (Greenberg, Demayo et al. 1995). This expression is not totally specific but specificity is significantly improved by the inclusion of MAR (matrix attachment regions) from the chick lysozyme gene (Greenberg, Demayo et al. 1994).
  • MAR matrix attachment regions
  • the 430 bp promoter region is strongly responsive to androgen induction and androgen response elements which bind the androgen receptor (AR) have been characterised (Claessens, Rushmere et al. 1990; Kasper, Rennie et al. 1994; Matusik, WO9403594; Rennie, Bruchovsky et al. 1993).
  • a negative regulatory region between bases -426 and -286 has also been identified (Rennie, Bruchovsky et al. 1993).
  • the PSA upstream region (to -630 bp) also acts as a strongly androgen responsive promoter and androgen response elements have also been characterised. This region is not sufficient to direct tissue specific expression in transgenic mice, however.
  • An enhancer region has recently been identified in the region 4 to 5kb upstream of the transcription start site.
  • the PSA enhancer has been shown to act as an androgen-inducible enhancer and in combination with the PSA promoter to display significant cell-type specificity (Henderson, W09519434; Schuur, Henderson et al. 1996). Also Pang et al. have reported that the equivalent promoter region isolated from a prostate cancer patient contained 7 mutations compared to the published sequence and was highly active in the prostate cancer cell line LNCaP (Belldegrun and Pang, W09614875; Pang, Taneja et al. 1995).
  • the viral SV40 enhancer was the first enhancer identified and it has been extensively characterised. It has the properties of enhancing expression from a variety of promoters whether it is upstream or downstream of the gene and is functional in either orientation. There are a limited number of reports where the SV40 enhancer has been used in conjunction with a steroid-responsive promoter. For example Israel and Kaufman (Israel and Kaufman 1989) reported substantial dexamethasone induction of expression (up to 17 fold) in constructs which combined the SV40 enhancer and adenovirus major late promoter with different copy numbers of glucocorticoid response elements derived from the mouse mammary tumour virus LTR promoter/enhancer. Wynshaw-Boris et al.
  • the glucocorticoid response region of the tyrosine amino transferase gene was able to confer 2 to 3 fold glucocorticoid ind icibility on the SV40 enhancer/early promoter (Grange, RO X et al. 1989). These examples indicate that continued steroid responsiveness is normally seen when the SV40 enhancer is present in conjunction with a steroid-responsive promoter.
  • the present inventors have combined, in various constructs, the viral SV40 enhancer with the promoter regions of two prostate-expressed, androgen-induced genes -probasin and PSA.
  • the promoter/enhancer combinations are thus now essentially androgen-independent, but maintain their specificity profile with respect to expression in prostate and non-prostate cells.
  • the probasin/SV40 enhancer combinations show svibstantial prostate specificity, while PSA/SV40 enhancer combinations are more promiscuous.
  • the present invention consists in a genetic construct providing preferential expression in specific tissue of a coding seqvience, the construct including the following elements (1) a steroid responsive promoter from a tissue specific gene, (2) a coding sequence, and (3) an SV40 enhancer, the elements being arranged as follows: the steroid responsive promoter from the tissLie specific gene being vipstream of the coding sequence and the SV40 enhancer being 3' of the coding sequence, or the SV40 enhancer being upstream of the coding sequence and the steroid responsive promoter from the tissue specific gene being positioned between the SV40 enhancer and the coding seqLience, or the steroid responsive promoter from the tissue specific gene being upstream of the coding sequence and the SV40 enhancer being positioned within an intron within the coding sequence; wherein the promoter has a lowered level of steroid responsiveness in the construct than in its native state.
  • the present invention consists in a genetic cassette comprising a steroid responsive promoter from a tissLie specific gene and an SV40 enhancer and an insertion site into which a coding sequence can be inserted, the insertion site being adjacent to and downstream of the promoter, wherein the promoter has a lowered level of steroid responsiveness in the cassette than in its native state.
  • the SV40 enhancer may be upstream of the tissue specific promoter or vice versa.
  • the promoter from the tissue specific gene is androgen-responsive. It is also preferred that the tissue specific gene is prostate specific.
  • tissue specific promoter is the probasin promoter, in particular Pb430 or Pb286 as described herein.
  • promoter is the PSA promoter, in particular PSA630 as described herein.
  • the present invention consists in a vector including the genetic construct of the first aspect of the present invention or the genetic cassette of the second aspect of the present invention. It is presently preferred that the vector is human adenovirus Type 5 or ovine adenovirus.
  • the present invention provides methods of treatment of cancers, in particular, prostate, bladder and breast, involving gene therapy using the constructs of the present invention.
  • cancers in particular, prostate, bladder and breast
  • the use of these constracts enables the use of androgen-ablation treatment in combination with the expression of therapeutic genes in gene therapy for androgen-independent or androgen-dependent prostate cancer.
  • FIGURE LEGENDS Figure 1A shows a linearised map of the plasmid pCATSAT.
  • H(Hind III), Sp(Sph I), P(Pst ⁇ ), S(SalI), X(Xba ⁇ ) and BamHI are shown; amp and ori are the plasmid ampicillin resistance gene and origin of replication respectively.
  • the chloramphenicol acetyl transferase (CAT) and serine acetyl transferase genes are indicated as is the rous sarcoma vims promoter (RSV) and polyadenylation regions derived from the hviman growth hormone gene or SV40.
  • Figure IB shows a niimber of promoter/enhancer constructs made in the pCATSAT vector.
  • Promoter regions Lipstream of the CAT gene were derived from either the probasin (Pb430 or Pb286) or PSA
  • the SV40 enhancer is shown as a dark box, its orientation being indicated by the position of the Ncol (N) site.
  • Figure 2 shows the sequence of SV40 enhancer.
  • the BstYI sites used for cloning and the unique Ncol site are underlined.
  • Figure 3 shows the transcriptional activity of constructs containing the Pb430 promoter in a number of cell types.
  • Figure 4 shows the transcriptional activity of constmcts containing the 286 base pair probasin promoter in a number of cell types.
  • Figure 5 shows the transcriptional activity of constracts containing the 630 base pair PSA promoter in a number of cell types.
  • Figure 6 shows the recombinant adenoviras Ad5SVbPb430PNP.
  • the cassette containing the SVbPb430 enhancer/promoter, the E.coli pLirine nucleoside phosphorylase gene (PNP) and SV40 polyadenylation region (SVpolyA) is inserted into the deleted Ela/6 region of the human adenovirus type 5.
  • PNP E.coli pLirine nucleoside phosphorylase gene
  • SVpolyA SV40 polyadenylation region
  • Figure 7A shows the effect of the combination of treatment with either Ad5SVbPb430PNP (SVPb) or Ad ⁇ PSAPNP in combination with 6-MPDR or PC3 cell viability. Viability was measured as metabolic activity relative to control, without virus or drag. MOI - multiplicity of infection.
  • Figure 7B shows the effect of virus phis drag treatment on MRC5 cells.
  • Figure 7C shows the effect of viras plus drug treatment on HepG2 cells.
  • plasmids combining the SV40 enhancer region in different positions and orientation with the promoter regions of the rat probasin and human PSA genes are diagrammed in Figure 1.
  • All plasmids have been prepared Lising the base plasmid pCATSAT.
  • This plasmid was derived from the plasmid pCAT-basic (Promega) by insertion into the BamHI site of a reference gene, serine acetyl transferase (SAT,)
  • SAT serine acetyl transferase
  • This reference reporter gene is under the control of the ubiqLiitously-expressed rous sarcoma virus promoter (RSV) and contains a 3' polyadenylation region derived from the human growth hormone gene.
  • RSV ubiqLiitously-expressed rous sarcoma virus promoter
  • a BamHI site was re- generated as shown between the SV40 and Imman growth hormone 3' polyadenylation regions.
  • Probasin sequences from -426 (Hindlll) or -286 to the Sad site at position + 28 were isolated by PCR from rat genomic DNA and cloned into pCATSAT to produce the Pb430 and Pb286 constructs, respectively.
  • the PCR fragment was cloned as a Sad fragment into the Sad site of pBluescriptSK+ using the Sad site at +28 and a site incorporated into the 5' primer. It was then recloned as a Hindlll to Xbal fragment into pCATSAT.
  • the Pb286 PCR fragment was cloned into pBluescriptSK+ as an EcoRI to Spel fragment using sites contained in the 5' and 3' primers respectively; it was then cloned as an Hindlll to Spel fragment into Hindlll-Xbal cut pCATSAT.
  • the PSA630 plasmid contained sequences 5' of the PSA transcription start site from EcoRI site (-630) to the Hindlll site (+6) which were isolated from human genomic DNA by PCR amplification.
  • SV40 enhancer sequences were isolated as a BstYI fragment from the plasmid pCAT-enhancer (Promega). The sequence of the enhancer region is shown in FigLire 2. The uniqLie Ncol site is underlined. This fragment was subcloned into the BamHI sites of the plasmids Pb430CATSAT and PSA630CATSAT to produce the plasmids Pb430CATSATSVa and . Pb430CATSATSVb and PSA630CATSATSVa and PSA630CATSATSVb respectively. The a and b orientations of the enhancer are distinguished by the position of the Ncol site as indicated in Figure 1.
  • the BstYI enhancer fragment was cloned into BamHI-cut pBluescriptSK+ (Stratagene), then re-cloned as a Xbal to Kpnl fragment into pUCl9. It was then excised as a Hindlll fragment and cloned into the Hindlll site of Pb430CATSAT to give the plasmids SVaPb430CATSAT and SVbPb430CATSAT.
  • the SV40 enhancer was cloned in front of the Pb286 promoter in the pBluescriptSK+ plasmid using BamHI site incorporated in the 5' PCR primer.
  • a range of cell types were transfected with the Pb430CATSAT construct and plasmids containing the SV40 enhancer in either orientation either downstream of the CAT gene or immediately upstream of the probasin promoter.
  • LNCaP LNCaP
  • PC-3 obtained from the ATCC
  • Tumour cell lines derived from other tissues included the breast cancer line MCF-7 (obtained from the ATTC), the liver cell line HepG2 cells (from Dr. G. Schreiber), and the bladder cancer cell line BL13 (Russell, Wass et al. 1989).
  • Human 293 embryonic kidney cells, adenovirus transformed, were obtained from Dr. F. Graham.
  • the normal lung fibroblast line MRC5 and the Chinese hamster ovary line CHO Kl were obtained from Dr. R. Holliday.
  • LNCaP cells were maintained in T-medium (Thalmann, Sikes et al. 1996).
  • PC-3, DU145 and TSLIP ⁇ MCF7 and BL13 cells were maintained in RPMI 1640 medium containing 2mM glutamine, non-essential amino acids and 10% fetal bovine seram (FBS).
  • HepG2 and MRC5 cells were maintained in Dulbecco's MEM containing 2 mM glutamine, 0.45% glucose and 10% FBS.
  • CHO and 293 cells were maintained in MEM containing non-essential amino acids and 10% FBS.
  • Transfections of DU145 and TsuPr cells were done Lising another cationic lipid transfection reagent CS06 and a similar protocol provided by Dr. T. Lockett (other transfection reagents such as Lipofectamine may also be Lised).
  • CAT and SAT assays At 44-48hr post-transfection cells were rinsed three times with PBS, and harvested by scraping into a 1.5ml centrifuge tube and brief centrifugation. Cell pellets were resuspended in 60 ⁇ l of 0.1M Tris.HCl, pH7.5, containing 500 ⁇ M Pefabloc protease inhibitor (Boehringer). Cells were lysed by three cycles of freezing and thawing and debris was pelleted for 5min in a microfuge. Supernatant (20 ⁇ l) was set aside for SAT assays and the remaining extract was heated at 65°C for 10 min. Debris was again pelleted and the supernatant used for CAT assays (Sleigh 1986).
  • Non-heated extract was used for assays of serine acetyl transferase activity.
  • Reaction mixtures (20 ⁇ l) comprised 2 or 4 ⁇ l of cell extract, ImM acetyl coenzyme A and 200uM serine containing O.l ⁇ Ci 14C-serine (Amersham, 50 mCi/ml, 150 mCi/mmol). Aliquots were removed between 20 and 120 min and reactions were stopped by heating at 95°C for 3 min. Reactions products were separated by thin layer chromatography and subjected to phosphor image analysis (Molecular Dynamics). The extent of conversion of serine to acetylated serine was determined using ImageQuant software. The ratio of RSVCAT to RSVSAT in the control transfection in each experiment was used to normalise promoter activities relative to the RSV promoter.
  • FigLire 2 Activities of the constructs containing the Pb430 promoter and different arrangements of the SV40 enhancer in different cell types are shown in FigLire 2.
  • the promoter alone shows low activity in the LNCaP prostate cell line and negligible activity in the PC-3 line which lacks an androgen receptor.
  • Co-transfection of the androgen receptor into PC-3 cells leads to a strong stimulation of expression, indicating the critical androgen- dependence of the promoter.
  • slight activity was seen in the MCF-7, breast, and HepG2, liver, cell lines.
  • DU145 and TsuPr prostate cell lines where expression ranged from 10 to 60% that of the RSV promoter. This high level expression is seen in the absence of co- transfection of the androgen receptor. Indeed, when the androgen receptor was co-transfected activity was generally found to be lower in PC-3 cells, perhaps reflecting competition between binding factors or stimulatory pathways. A similar effect was seen in the DU145 and TSLIP ⁇ lines (data not shown).
  • the Pb430SVa, SVaPb430 and SVbPb430 constracts showed 3.5 to 6 fold higher expression in the prostate PC-3 and DU145 cells than in any of the other cell types tested.
  • Tims the probasin promoter/SV40 enhancer combinations provide for substantial specificity for expression in the PC-3 androgen-independent prostate cells relative to other cell types.
  • the lack of reqLiirement for the presence of androgen receptor for high level expression also provides for a significant improvement in utility compared to the probasin promoter alone, as these combinations will be active in androgen-independent cancer cells and can be used while continuing androgen ablation therapy designed to eliminate androgen- dependent tumour cells.
  • a range of cell types were transfected with the Pb286CATSAT constract and plasmids containing the SV40 enhancer in either orientation either immediately upstream of the Pb286 promoter.
  • the Pb286 promoter is very poorly expressed in PC-3 cells, but is highly expressed when co-transfected with the androgen receptor.
  • combination of the Pb286 promoter with the SV40 enhancer are highly expressed in PC-3 cells in the absence of the androgen receptor.
  • For the SVb enhancer orientation expression was 70% of that seen in the presence of the androgen receptor, while for the SVa orientation expression was reduced in the presence of the AR (similarly to the Pb430 constracts). While expression was higher in PC-3 cells than in non-prostate cell types, the specificity of expression was less than that seen for the Pb430 constructs. The effect of the enhancer is thus similar to that seen for the Pb430 constracts in bypassing the requirement for androgen induction for expression from the promoter.
  • PSA630, PSA630SVa and PSA630SVb was examined in a number of cell lines following transfection alone or in combination with the AR gene. Results are shown in Figure 5. Expression from the PSA630 promoter was found to be significantly increased in the presence of AR in all cell types studied, though the level of indLiction varied considerably. Constructs containing the SV40 enhancer in either orientation 3' of the CAT gene showed substantially less enhancement of expression in the presence of AR in almost all cell types. The effect of AR ranged from three fold induction to two fold repression in different instances. The only exception was in MCF-7 cells where the AR enhanced expression by 5.8 and 4.3 fold for the SVa and SVb constructs respectively compared to tenfold in the absence of the enhancer. While these PSA630- based constracts show poor prostate specificity the effect of the SV40 enhancer on expression from this androgen-responsive promoter is similar to that on the Pb promoters in that expression becomes substantially androgen- independent.
  • the promoter cassette SVbPb430 was excised from pSVbPb430CATSAT as a Sail to Xbal fragment and blunt-ended. It was re- cloned in front of the E.coli pLirine nucleoside phosphorylase (PNP) gene in the plasmid pXCX3/PSAPNPase (Lockett et al. 1997) which had been cut with Kpnl and Hindlll, blunt-ended and re-ligated to remove the PSA promoter and subsequently cut with Spel and bkmt-ended.
  • PNP E.coli pLirine nucleoside phosphorylase
  • Ad ⁇ PSAPNP in which the expression of the PNP gene was under the control of the promoter of the prostate specific antigen (PSA) gene.
  • PSA prostate specific antigen
  • Ad5SVbPb430PNP is substantially more effective in killing the prostate PC-3 cells than Ad ⁇ PSAPNP; an equivalent suppression was seen at viras inputs at least 10 fold lower.
  • the same differential in killing was not seen for the non-prostate cells MRC-5 and HepG2; the two virases showed equivalent SLippression of HepG2 cells, while significant reduction in metabolic activity of MRC-5 cells following infection with Ad5SVbPb430PNP was only seen at the highest viras level tested (200 pfu).
  • Virus inputs required for equivalent cell killing were 5 to 10 fold higher for the non-prostate cells than for the PC-3 prostate cell line.
  • SVbPb430 promoter provides for substantially improved activity in prostate cells compared with the PSA promoter, and shows significant selectivity for prostate cells.
  • these promoter/enhancer combinations are new and have much greater utility than the original promoters in that they can be used to control expression of genes in a prostate-specific manner in both androgen-responsive and androgen- independent prostate cells and in the absence of androgens (as in cancer patients undergoing androgen-ablation treatment). This is of particular importance for example in the expression of therapeutic genes in gene therapy for androgen-independent prostate cancer.
  • constructs of the present invention will also be LiseiLil where patient has been treated surgically (castration) or with inhibitors of androgen synthesis or action.
  • SLich androgen- blockade therapy can be continLied without substantially diminishing the activity of these promoters.

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Abstract

L'invention concerne une construction génétique permettant l'expression préférentielle, dans un tissu spécifique, d'une séquence codante. La construction comprend les éléments suivants: (1) un promoteur sensible aux stéroïdes, provenant d'un gène spécifique de tissu, (2) une séquence codante, et (3) un activateur SV40. Ces éléments sont disposés de la façon suivante: le promoteur sensible aux stéroïdes provenant du gène spécifique de tissu est en amont de la séquence codante et l'activateur SV40 en position 3' de la séquence codante; l'activateur est en amont de la séquence codante, et le promoteur entre l'activateur et la séquence codante; ou bien le promoteur est en amont de la séquence codante et l'activateur à l'intérieur d'un intron de la séquence codante. La sensibilité du promoteur aux stéroïdes est plus faible dans la construction qu'à l'état natif.
PCT/AU1998/000057 1997-02-03 1998-02-03 Promoteurs specifiques de tissu WO1998033903A1 (fr)

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CA002279552A CA2279552A1 (fr) 1997-02-03 1998-02-03 Promoteurs specifiques de tissu
EP98901256A EP1009820A1 (fr) 1997-02-03 1998-02-03 Promoteurs specifiques de tissu
AU57421/98A AU5742198A (en) 1997-02-03 1998-02-03 Tissue specific promoters

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AUPO4923 1997-02-03
AUPO4923A AUPO492397A0 (en) 1997-02-03 1997-02-03 Prostate specific promoters

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JP2002537807A (ja) * 1999-03-01 2002-11-12 コモンウェルス サイエンティフィック アンド インダストリアル リサーチ オーガニゼーション 前立腺特異的膜抗原遺伝子のイントロン３を含む調節構築物
US7723104B2 (en) 2004-04-02 2010-05-25 Board Of Regents, The University Of Texas System Cancer specific promoters
US8569051B2 (en) 2008-02-14 2013-10-29 Mogam Biotechnology Research Institute Expression vector for gene therapy

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CN102628041B (zh) * 2008-02-14 2013-09-11 财团法人牧岩生命工学研究所 适于表达用于基因治疗的编码序列的表达载体
CN102618536B (zh) * 2008-02-14 2013-10-16 财团法人牧岩生命工学研究所 适于表达用于基因治疗的编码序列的表达载体
CN102634515B (zh) * 2008-02-14 2013-11-06 财团法人牧岩生命工学研究所 适于表达用于基因治疗的编码序列的表达载体
CN109988759A (zh) * 2017-12-29 2019-07-09 上海细胞治疗研究院 一种在t细胞中具有高转录活性的嵌合启动子

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BRITISH JOURNAL OF UROLOGY, (March 1997), Volume 79, Supplement 1, DANNULL J. and BELLDEGRUM A.S., "Development of Gene Therapy for Prostate Cancer Using a Novel Promoter of Prostate Specific Antigen", pages 97-103. *
CANCER RESEARCH, (1 February 1997), Volume 57, PANG S. et al., "Identification of a Positive Regulatory Element Responsible for Tissue-Specific Expression of Prostate-Specific Antigen", pages 495-499. *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002537807A (ja) * 1999-03-01 2002-11-12 コモンウェルス サイエンティフィック アンド インダストリアル リサーチ オーガニゼーション 前立腺特異的膜抗原遺伝子のイントロン３を含む調節構築物
EP1157105A4 (fr) * 1999-03-01 2003-01-15 Commw Scient Ind Res Org Produits de recombinaison regulateurs comprenant l'intron 3 du gene de l'antigene d'enveloppe prostatique specifique
US7074400B1 (en) 1999-03-01 2006-07-11 The Commonwealth Of Australia Regulatory constructs comprising intron 3 of prostate specific membrane antigen gene
US7871819B2 (en) 1999-03-01 2011-01-18 Commonwealth Scientific And Industrial Research Organisation Regulatory constructs comprising intron 3 of prostate specific membrane antigen gene
US7723104B2 (en) 2004-04-02 2010-05-25 Board Of Regents, The University Of Texas System Cancer specific promoters
US7816131B2 (en) 2004-04-02 2010-10-19 Board Of Regents, The University Of Texas System Cancer specific promoters
US8569051B2 (en) 2008-02-14 2013-10-29 Mogam Biotechnology Research Institute Expression vector for gene therapy
US8617875B2 (en) 2008-02-14 2013-12-31 Mogam Biotechnology Research Institute Expression vector suitable for expression of a coding sequence for gene therapy
US8628956B2 (en) 2008-02-14 2014-01-14 Mogam Biotechnology Research Institute Expression vector suitable for expression of a coding sequence for gene therapy

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CN1250477A (zh) 2000-04-12
EP1009820A1 (fr) 2000-06-21
AUPO492397A0 (en) 1997-02-27
CA2279552A1 (fr) 1998-08-06

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