[go: up one dir, main page]

WO2017036052A1 - Promoteur recombinant régulé par l'acide rétinoïque avec un effet dépendant de la concentration et utilisation correspondante - Google Patents

Promoteur recombinant régulé par l'acide rétinoïque avec un effet dépendant de la concentration et utilisation correspondante Download PDF

Info

Publication number
WO2017036052A1
WO2017036052A1 PCT/CN2015/100359 CN2015100359W WO2017036052A1 WO 2017036052 A1 WO2017036052 A1 WO 2017036052A1 CN 2015100359 W CN2015100359 W CN 2015100359W WO 2017036052 A1 WO2017036052 A1 WO 2017036052A1
Authority
WO
WIPO (PCT)
Prior art keywords
retinoic acid
gene
promoter
expression
concentration
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2015/100359
Other languages
English (en)
Chinese (zh)
Inventor
李小彦
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Publication of WO2017036052A1 publication Critical patent/WO2017036052A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • 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
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids

Definitions

  • the invention belongs to the field of medical biotechnology research and relates to a recombinant promoter with concentration-dependent effect regulated by retinoic acid.
  • the promoter is composed of a regulatory box A and a control box B, and the control box A comprises 10 DR5 elements. (5'-GTTCAC-3') is composed in series, and the control box B is a 115 bp artificially designed assisted starting element.
  • the reporter gene linked to the recombinant promoter can be used to display and detect the concentration level of retinoids in the extracellular fluid or environment; the recombinant promoter can be used to regulate the expression of the gene of interest in cells or animals in vitro; Recombinant promoters can also be used to regulate the expression of therapeutic genes in human gene therapy.
  • a retinoic acid response element exists in the genome of a natural organism, and the retinoic acid response element is generally present in the upstream sequence of the gene regulatory promoter in two to four repeating arrangements.
  • the retinoid response element in conjunction with other transcription factor regulatory elements, collectively regulates the transcriptional expression of downstream genes.
  • the base sequence composition of the retinoic acid response element generally has multiple types, which are named DR1, DR2, DR3, DR4 and DR5 elements, respectively.
  • RAR and RXR form a heterodimer, they recognize the retinoic acid response element on the genome and bind to it.
  • the retinoic acid enters the nucleus and binds to RAR/RXR. The expression of downstream genes is initiated.
  • the retinoic acid-RAR/RXR needs to work in concert with other transcription factors (such as NF- ⁇ B or STAT) to initiate downstream genes. expression.
  • retinoic acid-regulated promoter there is no concentration-dependent retinoic acid-regulated promoter in nature.
  • Previous researchers have attempted to construct a retinoic acid-regulated promoter with a concentration-dependent effect, but they are not perfect.
  • Jason Aoto et al.1 constructed a retinoic acid-regulated promoter consisting of 6 DR5 elements and a thymidine.
  • the kinase promoter (Thymidine Kinase, TK promoter, 750 bp) is composed.
  • Figure 3C published by the author, it is clear that EGFP still has a distinct background expression in the absence of retinoic acid. .
  • thymidine kinase promoter as a co-promoter element, and the thymidine kinase promoter itself has independent transcriptional activity in retinoic acid. In the absence of -RAR/RXR, the expression of downstream genes can still be initiated.
  • a retinoic acid-regulated promoter with a concentration-dependent effect was invented (ie, when the retinoic acid was not added, the downstream gene was not expressed, and as the concentration of retinoic acid increased, the downstream gene gradually increased the expression level), not only for in vitro experimental studies. It is a good gene regulation tool, and it is also a new therapeutic gene regulation method for gene therapy in vivo.
  • the present invention is based on the recombination of DR5 elements, using a series of 10 DR5 elements in series to form a control box A, the base sequence of which is as follows:
  • the control box B is a 115 bp artificially designed assisted activation element whose base sequence is as follows:
  • the complete base sequence of the promoter is as follows:
  • the invention also discloses a method for applying a recombinant promoter having a concentration-dependent effect regulated by retinoic acid.
  • the promoter-linked green fluorescent protein or luciferase reporter gene has a method for detecting and detecting trace levels of retinoic acid in an extracellular fluid or environment, and is a method for detecting a retinoid.
  • a retinoid drug such as all-trans retinoic acid
  • a retinoid drug is added to regulate the expression of the target gene, which is used to regulate the expression of the target gene in cells or animals in vitro.
  • intravenous or oral retinoids (such as all-trans retinoic acid) regulate the expression of therapeutic genes, which are used to regulate therapeutic gene expression in human gene therapy.
  • the present invention also indicates that the promoter can be used to regulate the expression of the T cell chimeric antigen receptor (CAR-T) gene and avoid the formation of cytokine storms.
  • CAR-T T cell chimeric antigen receptor

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Analytical Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Plant Pathology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne un promoteur recombinant, dont la longueur de séquence entière est de 297 pb, constitué d'une boîte de régulation A et d'une boîte de régulation B. La boîte de régulation A contient 10 éléments DR5 (5'-GTTCAC-3') en série et la boîte de régulation B est un élément de départ d'assistance de conception artificielle de 115 pb. Le promoteur recombinant est régulé par l'acide rétinoïque dans le fluide extracellulaire et la quantité d'expression de la protéine cible régulée par celui-ci a un effet dépendant de la concentration en acide rétinoïque.
PCT/CN2015/100359 2015-08-30 2015-12-31 Promoteur recombinant régulé par l'acide rétinoïque avec un effet dépendant de la concentration et utilisation correspondante Ceased WO2017036052A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201510539568.5A CN106480024B (zh) 2015-08-30 2015-08-30 受维甲酸调控的具有浓度依赖效应的重组启动子及其使用
CN201510539568.5 2015-08-30

Publications (1)

Publication Number Publication Date
WO2017036052A1 true WO2017036052A1 (fr) 2017-03-09

Family

ID=58186535

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2015/100359 Ceased WO2017036052A1 (fr) 2015-08-30 2015-12-31 Promoteur recombinant régulé par l'acide rétinoïque avec un effet dépendant de la concentration et utilisation correspondante

Country Status (2)

Country Link
CN (1) CN106480024B (fr)
WO (1) WO2017036052A1 (fr)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991007488A1 (fr) * 1989-11-16 1991-05-30 The Salk Institute For Biological Studies Elements de reponse de l'acide retinoique beta; compositions et analyses
WO1992016658A1 (fr) * 1991-03-18 1992-10-01 The Salk Institute For Biological Studies Compositions a elements de reaction et dosages les incorporant

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5922596A (en) * 1990-12-10 1999-07-13 Bristol-Myers Squibb Co. Promoter of the retinoic acid receptor gene for directing gene expression
FR2732035B1 (fr) * 1995-03-23 1997-05-30 Agronomique Inst Nat Rech Procede de regulation de l'expression d'un gene dans un baculovirus, par un site de fixation d'un recepteur de l'acide retinoique, et vecteur pour la mise en oeuvre du dit procede
WO2001016368A1 (fr) * 1999-09-01 2001-03-08 Bristol-Myers Squibb Company Systemes de transcription in vitro et ses utilisations

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991007488A1 (fr) * 1989-11-16 1991-05-30 The Salk Institute For Biological Studies Elements de reponse de l'acide retinoique beta; compositions et analyses
WO1992016658A1 (fr) * 1991-03-18 1992-10-01 The Salk Institute For Biological Studies Compositions a elements de reaction et dosages les incorporant

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
AOTO, J. ET AL.: "Synaptic Signaling by All-Trans Retinoic Acid in Homeostatic Synaptic Plasticity", NEURON, vol. 60, no. 2, 23 October 2008 (2008-10-23), pages 308 - 320, XP055365143, ISSN: 0896-6273 *
SUN, YUQIANG ET AL.: "Advanced Progress on The Relationship Between RA and Its Receptors and Malignant Tumors", CHINESE BULLETIN OF LIFE SCIENCE, vol. 26, no. 3, 15 March 2014 (2014-03-15), pages 295 - 301, ISSN: 1004-0374 *

Also Published As

Publication number Publication date
CN106480024A (zh) 2017-03-08
CN106480024B (zh) 2020-10-20

Similar Documents

Publication Publication Date Title
Gigueère Retinoic acid receptors and cellular retinoid binding proteins: complex interplay in retinoid signaling
Grausenburger et al. Conditional deletion of histone deacetylase 1 in T cells leads to enhanced airway inflammation and increased Th2 cytokine production
Komuczki et al. Fate-mapping of GM-CSF expression identifies a discrete subset of inflammation-driving T helper cells regulated by cytokines IL-23 and IL-1β
Gao et al. Understanding the role of Tbx1 as a candidate gene for 22q11. 2 deletion syndrome
Liu et al. Emerging roles and mechanisms of long noncoding RNAs in atherosclerosis
Lotan Suppression of squamous cell carcinoma growth and differentiation by retinoids
Hu et al. MicroRNA-155 confers encephalogenic potential to Th17 cells by promoting effector gene expression
Sheikh et al. Cutting edge: IFN-γ is a negative regulator of IL-23 in murine macrophages and experimental colitis
Roberts Regulating retinoic acid availability during development and regeneration: the role of the CYP26 enzymes
Satoh et al. The spatial patterning of mouse cone opsin expression is regulated by bone morphogenetic protein signaling through downstream effector COUP-TF nuclear receptors
JP2013535969A5 (fr)
Fan et al. Transient, inducible, placenta-specific gene expression in mice
JP6022470B2 (ja) 分散型タンパク質形質導入ドメインを含む誘導性発現系転写モジュレータおよびその使用方法
Tofukuji et al. Identification of a new pathway for Th1 cell development induced by cooperative stimulation with IL-4 and TGF-β
Noel et al. KEAP1 editing using CRISPR/Cas9 for therapeutic NRF2 activation in primary human T lymphocytes
Crowe Retinoic acid mediates post-transcriptional regulation of keratin 19 mRNA levels
Li et al. Targeted overexpression of prostacyclin synthase inhibits lung tumor progression by recruiting CD4+ T lymphocytes in tumors that express MHC class II
Jia et al. Innate lymphoid cells: a new key player in atopic dermatitis
CN112553196B (zh) 核酸序列及其作为启动子的应用
Wang et al. Functional impact of heterogeneous nuclear ribonucleoprotein A2/B1 in smooth muscle differentiation from stem cells and embryonic arteriogenesis
Saleh et al. STAT3/SOCS3 axis contributes to the outcome of salmonid whirling disease
WO2017036052A1 (fr) Promoteur recombinant régulé par l'acide rétinoïque avec un effet dépendant de la concentration et utilisation correspondante
Glenn et al. Control of rene gene expression
Wang et al. Gene regulation of CYP4F11 in human keratinocyte HaCaT cells
Kwon et al. Foxp3 induces IL-4 gene silencing by affecting nuclear translocation of NFκB and chromatin structure

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: 15902856

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: 15902856

Country of ref document: EP

Kind code of ref document: A1