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WO1999057265A1 - Arn de leurrage chimeres ayant une action anti-vih synergique - Google Patents

Arn de leurrage chimeres ayant une action anti-vih synergique Download PDF

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Publication number
WO1999057265A1
WO1999057265A1 PCT/US1998/004219 US9804219W WO9957265A1 WO 1999057265 A1 WO1999057265 A1 WO 1999057265A1 US 9804219 W US9804219 W US 9804219W WO 9957265 A1 WO9957265 A1 WO 9957265A1
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Prior art keywords
tar
rev
elements
hiv
group
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PCT/US1998/004219
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English (en)
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Julianna Lisziewicz
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Julianna Lisziewicz
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Priority to PCT/US1998/004219 priority Critical patent/WO1999057265A1/fr
Priority to AU91967/98A priority patent/AU9196798A/en
Publication of WO1999057265A1 publication Critical patent/WO1999057265A1/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
    • 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
    • C12N15/1131—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 against viruses
    • C12N15/1132—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 against viruses against retroviridae, e.g. HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/11—Antisense
    • C12N2310/111—Antisense spanning the whole gene, or a large part of it
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/13—Decoys
    • 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
    • C12N2799/00—Uses of viruses
    • C12N2799/02—Uses of viruses as vector
    • C12N2799/021—Uses of viruses as vector for the expression of a heterologous nucleic acid
    • C12N2799/027—Uses of viruses as vector for the expression of a heterologous nucleic acid where the vector is derived from a retrovirus

Definitions

  • the present invention relates generally to the field of treatment of viral diseases in human beings and animals. More specifically, it relates to materials and methods for intracellular immunization against Human Immunodeficiency Virus (HIV) infection.
  • HIV Human Immunodeficiency Virus
  • the preferred inhibitory construct is the combination of Tat-binding and Rev-binding RNA, preferably expressed together from an HIV-inducible promoter. This Tat-binding and Rev-binding chimeric RNA is the first drug which has a strong synergistic antiviral activity among the domains of a single molecule.
  • the tat protein of HIV-1 transactivates viral gene expression and is essential for viral replication.
  • the tat activation response element (TAR) has been localized within the region of the first 44 nucleotides downstream of the transcription initiation site. This region, present in all HIV-1 transcripts, forms an unusually stable stem loop structure, and several lines of evidence suggest that the transcriptional effect of the tat protein is mediated through its interaction with the TAR region of viral RNA. It has been demonstrated that the Tat protein binds to the TAR RNA sequences.
  • Rev protein of HIV-1 is another important factor in viral replication. It 2 helps activate HIV-1 gene expression, and part of its mechanism for action involves binding with a portion of the HIV RNA.
  • Rev response element the Rev response element (RRE) (Malim et al.; HIV-1 structural gene expression requires binding of the Rev transactivator to its RNA target sequence. Cell 60:675-683 (1990).
  • the Rev protein acts after the transcription phase to facilitate both the transport of incompletely spliced viral mRNA from the infected cell's nucleous to the cell's cytoplasm and the production of viral structural proteins (Malim et al.; the HIV-1 Rev transactivator acts through a structured target sequence to activate nuclear export of unspliced viral mRNA 1989, Nature 338:254- 257; Hope et al.; Trans-dominant inhibition of HIV-1 Rev occurs through formation of inactive protein complexes 1992 J. Virol. 66:1849-1855).
  • Rev proteins bind to the RRE RNA element and the Rev proteins together with other cellular proteins can transport unspliced and singly spliced HIV-1 RNA from the nucleus to the cytoplasm (Meyer et al.; The HIV-1 Rev transactivator shuttles between the nucleus and the cytoplasm, 1994 Genes Dev. 8:1538-1547).
  • RNA is ready to be packaged into a protective envelope, or capsid, and the resulting infectious viral particles, or virions, can then be released from the cell.
  • Polymeric-TAR genes Genetic constructs using multiple copies of DNA that encode TAR RNA (Polymeric-TAR genes) have been developed and explored for their ability to block Tat protein activity. Polymeric TAR was combined with an antisense-faf gene (J. Lisziewicz et al.; An autoregulated dual-function antitat gene for HIV-1 gene therapy. J. Virol. 69:1 206-12 (1995); J. Lisziewicz et al; Antitat gene therapy: a candidate for late-stage AIDS patients.
  • the subject invention concerns materials and methods to inhibit viral replication and to treat viral infections in humans and animals.
  • the materials and methods of the subject invention are described in terms of lentiviruses, but one of ordinary skill in the art will recognize they are applicable to a variety of viruses.
  • the subject invention pertains to new RNA molecules comprised of selected RNA elements, preferably including one or more TAR element transcribed in tandem with one or more Rev binding elements transcribed in tandem, more preferably including 2-7 TAR elements and 2-4 Rev binding elements, and most preferably including 5 TAR elements and 2 Rev binding elements.
  • Rev binding elements are known in several forms, and preferred forms include the wild-type RRE, Aptamer 1 (Apt1), Aptamer 2(Apt2), bul and 2 or more bul (2bul, 3bul, etc.).
  • Another embodiment of the present invention is a DNA construct having a vector and a promoter operably linked to a unit having at least two to seven, more preferably five DNA segments encoding HIV-1 Tat activation response (TAR) elements so that the TAR elements are transcribed in tandem, and at least one, more preferably at least two, DNA segments encoding HIV-1 Rev binding elements so that the Rev binding elements are transcribed in tandem.
  • the DNA construct preferably includes a promoter capable of being regulated by HIV-1 Tat protein, and is preferably a primate lentivirus long terminal repeat (LTR) promoter such as the LTR promoter for Human Immunodeficiency Virus -1 "(HIV-1) LTR".
  • LTR primate lentivirus long terminal repeat
  • the constructs have two to five, preferably three, repeating units.
  • the present invention relates to articles including the RNA or DNA constructs and a delivery system.
  • the preferred delivery systems include viral gene delivery vectors, including adenovirus, herpesvirus, adeno-associated virus, SV-40 virus, retrovirus and lentivirus vectors as well as non-viral gene delivery systems, including liposomes, virosomes, and polyethylenimine conjugates and other conjugates.
  • non-viral gene delivery systems virosomes are preferred.
  • retrovirus or lentivirus vectors capable of integrating the construct into the genetic material in the nucleous of the target cell. Where retrovirus or lentivirus vectors are used, it is preferable to modify the cells in a transient fashion by loading them with dNTP. (See USSN 08/989,301 titled Materials and Methods for Gene Transfer, filed December 11 , 1997), in order to be able to transduce, that is, add the foreign genetic material to the quiescent (non- dividing) cells.
  • the preferred delivery systems include viral gene delivery systems and non-viral gene delivery systems.
  • the genetic constructs of the present invention are preferably delivered to cells using high efficiency transfection techniques such as antibody-mediated delivery of genes to cells.
  • a gene delivery complex compatible with a specific type of targeted cell is formed from a carrier, a delivery particle, and the DNA construct. Suitable materials and techniques are described in USSN
  • the subject invention also concerns a method for inhibiting viral replication which comprises administering a combination of multiple-TAR and multiple-REV binding elements, which the inventors have demonstrated to be advantageously included in a single molecule. These multiple elements can be administered in the form of a single molecule which demonstrates an enhanced ability to inhibit viruses dependent upon the activities of Tat and Rev proteins.
  • the genetic construct of the subject invention preferably also includes an inducible promoter to direct the expression of the TAR and Rev binding elements. Suitable promoters include for example, an HIV-1 -LTR promoter and modifications thereof. These constructs would work if they are expressed from constitutive promoters such as cytomegalovirus, SV40 promoter and Moloney murine leukemia virus LTR. However, inducible production of the product from the construct is advantageous as it limits the activity of the construct to a time and place where it is needed, thereby minimizing potential side effects.
  • An advantage of the present invention is that the DNA constructs which combine TAR and Rev binding elements as described herein yield greatly enhanced activity over similar constructs using the elements separately. Another advantage of the present invention is that it allows lower concentrations of the constructs to be used. Yet another advantage is that the constructs are active only in infected cells, and remain quiescent in uninfected cells. As a result, the potential side effects are minimized. Further, the combination of multiple TAR and RRE elements in the same antiviral vector targets two important viral proteins at the same time. This provides further constraints for any escape mutants, as such a mutant has to confer simultaneous mutations in both Rev and Tat proteins. Furthermore, this fusion molecule suppresses both virus replication and virus expression from infected cells.
  • Fig. 1 is a schematic representation of the polymeric-TAR and Rev binding
  • DNA constructs each of which has a promoter (LTR) and variable amounts of TAR and Rev binding elements.
  • LTR promoter
  • Fig. 2 depicts the known structure of the wild-type HIV-1 RRE and its variants aptamer 1 (Apt1), aptamer 2 (Apt2) and bul.
  • Fig. 3 is a comparison of anti-Rev activity of several different constructs, each having an LTR promoter and 5 TAR elements, with several Rev binding variants, including the wild-type RRE (wtRRE), Apt1 , Apt2 and bul.
  • wtRRE wild-type RRE
  • Apt1 Apt2
  • bul is shown as both 1 and 2 elements (bul and 2bul).
  • Fig. 4 is a comparison of anti-TAT activity of various DNA constructs, each of which has a promoter and 5 TAR elements, and variable types of Rev binding elements, shown as the Apt1 and 2bul variants.
  • Fig. 1 wild-type HIV-1 RRE
  • Fig. 3 is a comparison of anti-Rev activity of several different constructs, each having an LTR promoter and 5 TAR
  • LTR- OTAR is a control which is effectively the promoter only, and which does not have antiviral activity
  • LTR-4TAR is in prior art
  • the Rev binding elements are shown as 2bul and Apt1 variants.
  • Fig. 6 compares the anti-HIV activity of three different DNA constructs at varying concentrations. LTR-4TAR is in the prior art. Variable amounts of TAR are shown with one Rev binding element identified as the Apt1 variant.
  • Fig. 7 compares the anti-HIV activity of three different DNA constructs at varying concentrations.
  • LTR-4TAR is the prior art. Variable amounts of TAR are shown with two copies of the bul variant of the Rev binding element.
  • Fig. 8 depicts the mechanism of action of the constructs of the present invention.
  • Fig. 9 a schematic representation of the multi-unit polymeric-TAR and Rev binding constructs. Two examples are shown, for a non-viral gene delivery system (Plasmid constructs) and for a viral gene delivery system (Retrovirus vector construct).
  • the subject invention provides materials and methods to inhibit viral replication in humans and animals. Specifically provided herein are unique genetic constructs and gene therapy methods to inhibit or prevent infection by lentiviruses, including HIV. 9
  • Fig. 1 is a schematic representation of various polymeric-TAR and Rev binding constructs studied by the inventor. All constructs were made using the procedure described in Example 1 , below. All constructs contain a CD7-LTR element as a promoter, which is derived from the wild type HIV-1 -LTR, by deletion of the negative regulatory element (NRE). A plasmid construct, LTR-5TAR-CAT, was used as the backbone to extend the LTR-5TAR gene sequence with the previously described Rev binding elements.
  • Tandem bul termed 2bul, were obtained by ligating a second bul PCR product 3' of LTR-5TAR-bul. Plasmids pLTR-5TAR-Apt1 and pLTR-5TAR-2bul were treated with Bglll and relegated to obtain pLTR-1TAR-Apt1 and pLTR-1TAR-2bul constructs.
  • Fig. 2 depicts the known structure of the HIV-1 RRE and its variants Apt1 , Apt2 and bul.
  • Apt 1 and 2 has been selected because they had been reported to bind more strongly to the Rev protein in vitro than the wild type RRE.
  • Fig. 3. reports Anti-Rev activity of constructs having both TAR and Rev binding elements as percent Rev inhibition.
  • a cell line was altered to produce an easily measured protein (CAT) in response to the presence of the Rev protein.
  • CAT easily measured protein
  • a human kidney cell line 293T cells was used for the transient transfection experiment. Transfection efficiency as high as nearly 90% was achieved using the following Ca 3 (PO 4 ) 2 -based protocol with the 293T cells.
  • One microgram of DNA was used for each well of a 24-well plate.
  • the values as shown are the averages of three experiments. 50 ng pDM128 and 4 ng pCMV-Rev plasmid DNA was used to induce Rev-dependent CAT expression (Huang et al; J. Virol 65:2131-4 (1991)), and with 0.4 ng LTR-Tat to induce the expression of TAR-RRE genes. LTR-OTAR and carrier plasmid Bluescript was added so that the total amount of HIV-1 LTR was 200 ng in each transfection. LTR-OTAR was used as control. Forty-eight hours after transfection, the same number of cells were lysed and CAT protein was measured by ELISA according to the manufacturer's protocol (Boehringer Mannheim, Indianapolis, IN). The values as shown are the averages of three separate experiments.
  • Fig. 4 compares the anti-Tat activity of 5TAR, 5TAR-Apt1 , and 5TAR-2bul constructs.
  • LTR-CAT was cotransfected with LTR-Tat as described above at Fig. 3, and each of the antiviral genes as described earlier (Lisziewicz, J., et al.: Tat regulated production of multmerized TAR RNA inhibits HIV-1 gene expression. New Biologist 3: 82-90, 1991).
  • LTR-OTAR was used as a control. Forty-eight hours after transfection, the same number of cells were lysed and CAT protein was measured by ELISA according to the manufacturer's protocol (Boehringer Mannheim, Indianapolis, IN). Values are the averages of three separate experiments. This figure demonstrates that the most effective Rev binding elements (Apt1 and 2bul) in the combination molecule does not degrade the anti-Tat activity of the polymeric-TAR molecule.
  • Fig. 5 demonstrates the anti-HIV-1 activity of the different TAR and Rev binding constructs.
  • different antiviral vectors were cotransfected with pLW-int6, which encoded an integrase defective variant of HIV-1. The transient transfection was performed as described above at Fig. 3. At 72 hours post-transfection, p24 was measured from the cell culture supernatant using an ELISA system provided by Coulter. 12
  • LTR-OTAR is the control which does not have an antiviral activity.
  • the polymeric-TAR construct, LTR-4TAR is a prior art construct known to sequester the Tat protein, and it is used in this assay to demonstrate the state of art of the polymeric-TAR technology.
  • New constructs expressing Rev-binding motifs from HIV-1 LTR, 2bul and Apt1 have been shown in Fig. 3 to sequester Rev and therefore, as demonstrated in this figure, can inhibit HIV-1 production.
  • This figure also clearly demonstrates the superior anti-HIV activity of the combination constructs LTR-5TAR-2bul and LTR-5TAR-Apt1.
  • Fig. 6 demonstrates the anti-HIV-1 activity of the Apt1 Rev binding element alone and in combination with the polymeric TAR element. This experiment was done as described in Fig. 5 using LTR-OTAR as a control. Different amounts of LTR-5TAR-Apt1 plasmids were used in cotransfection to assess anti-HIV activity. Inhibition of HIV-1 replication is dependent on the concentration of the antiviral molecules which is also depicted on the figure by the trendlines of the different antiviral molecules. These trendlines describe that antiviral activity islogarithmicallyy increasing with the concentration of the antiviral RNA product, (or, drug). Mathematical analysis of the trendlines resulted in the following logarithmically equation between the concentration of the antiviral molecules (x) and % of HIV-1 inhibition (y).
  • Fig. 7. demonstrates the anti-HIV-1 activity of the 2bul Rev binding element alone and in combination with the polymeric TAR element. T;his experiment was done as described in Fig. 5 using LTR-OTAR as a control. Different amounts of LTR-5TAR-2bul plasmids were used in cotransfection to assess anti-HIV activity. Inhibition of HIV-1 replication is dependent on the concentration of the antiviral molecules which is also depicted on the figure by the trendlines of the different antiviral molecules. These trendlines describe that antiviral activity increases logarithmically with the concentration of the antiviral RNA (or, drug). Mathematical analysis of the trendlines resulted in the following logarithmically equation between the concentration of the antiviral molecules (x) and % of HIV-1 inhibition (y).
  • I (CX,1 in combination/CX,1) + (CX,2 in combination/CX,2) where CX,1 in combination and CX,2 in combination are the concentrations of elements 1 and 2, respectively, in the combined construct required to reduce HIV-1 production by X%. CX,1 and CX,2 are the concentrations of non-combination constructs required to achieve the same X% decrease.
  • 1 1 ; for synergy, I ⁇ 1 ; for antagonism, I >1.
  • I is computationally equivalent to the "combination index" introduced for mutually exclusive interaction in the "median effect analysis” (Chou, T.C., and Talalay, P.
  • Fig. 8. demonstrates the mechanism of action for the constructs of the present invention.
  • the TAR-RRE that is, polymeric- TAR and one or more Rev binding elements
  • the figure depicts an HIV-1 infected cell (see integrated provirus, HIV-1).
  • the Tat protein is produced by the provirus and activates the expression of HIV-1 LTR.
  • the polymeric-TAR-RRE RNA (the drug) is produced in the nucleus and the polymeric-TAR part of the molecule sequesters the Tat protein and the polymeric-RRE part of the molecule sequesters the Rev protein.
  • Sequestering Tat results in inhibition of the expression of all HIV-1 genes (including the rev gene).
  • Sequestering the Rev protein results in inhibition of the transport of single spliced and unspliced HIV-1 RNA from the nucleus to the cytoplasm. Therefore, both gene expression and provirus production is blocked in cells containing the chimeric antiviral gene.
  • Fig. 9 is a schematic representation of the multi-unit constructs.
  • Non viral gene delivery systems may use the plasmid construct.
  • This specific construct contains 3 copies of the TAR-RRE chimeric molecule, however depending on the size limitations, two and even one copy of the chimeric molecule can be used. If there is no size limitation on the gene delivery system, the use of higher numbers of units of the chimeric molecule is preferred, because, theoretically, maximal inhibition of HIV production will result. However, practically we expect complete inhibition of HIV with 3 units of the chimeric TAR-RRE because the strong synergistic antiviral activity of these molecules.
  • the other example is the retrovirus vector containing the 3x(5TAR-RRE) antiviral gene. Retrovirus vectors (including lentivirus vectors) are preferred gene delivery vehicles for these constructs because they can efficiently integrate the antiviral gene into the chromosome of the target 18

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Abstract

Cette invention concerne une construction comportant une unité qui comprend, d'une part, un premier groupe comprenant au moins une unité de l'élément de réponse d'activation de Tat (TAR) et, d'autre part, un second groupe comprenant au moins une unité d'un élément de liaison de Rev. Cette construction est utile afin de supprimer la production du virus VIH dans les cellules. La construction inhibitrice préférée consiste en une combinaison d'ARN liant le Tat et liant le Rev qui sont de préférence exprimés ensemble à partir d'un promoteur pouvant être induit par le VIH. Cet ARN chimère de liaison du Tat et de liaison du Rev représente le premier médicament à posséder une forte action antivirale synergique dans les domaines d'une seule molécule.
PCT/US1998/004219 1998-05-04 1998-05-04 Arn de leurrage chimeres ayant une action anti-vih synergique WO1999057265A1 (fr)

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PCT/US1998/004219 WO1999057265A1 (fr) 1998-05-04 1998-05-04 Arn de leurrage chimeres ayant une action anti-vih synergique
AU91967/98A AU9196798A (en) 1998-05-04 1998-05-04 Chimeric decoy rnas having synergistic anti-hiv activity

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002101036A1 (fr) * 2001-05-11 2002-12-19 Kazunari Taira Methode de formation d'un complexe stable d'un transcrit d'adn codant un peptide arbitraire a l'aide d'un produit de traduction, construction d'acide nucleique utilisee dans cette methode, complexe obtenu par cette methode et criblage de proteine fonctionnelle et marn ou adn codant la proteine a l'aide de cette methode
WO2016142948A1 (fr) 2015-03-11 2016-09-15 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Oligonucléotides leurres pour le traitement de maladies

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WO1993011230A1 (fr) * 1991-12-02 1993-06-10 Dynal As Cellule souche modifiee de mammifere bloquant la replication virale
WO1995031477A1 (fr) * 1994-05-18 1995-11-23 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Vecteurs a sequences tar multiples du vih-1 inhibant l'expression genique du vih-1

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WO1991010453A1 (fr) * 1990-01-18 1991-07-25 The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce Vecteur a elements multiples de reponse de cibles affectant l'expression de genes
WO1991014436A1 (fr) * 1990-03-21 1991-10-03 Isis Pharmaceuticals, Inc. Reactifs et procedes de modulation de l'expression de genes par homotypie d'arn
WO1993011230A1 (fr) * 1991-12-02 1993-06-10 Dynal As Cellule souche modifiee de mammifere bloquant la replication virale
WO1995031477A1 (fr) * 1994-05-18 1995-11-23 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Vecteurs a sequences tar multiples du vih-1 inhibant l'expression genique du vih-1

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Title
LEE, S.-W. ET AL.: "Inhibition of human immunodeficiency virus type 1 in human T cells by a potent rev response element decoy consisting of the 13-nucleotide minimal rev-binding domain", JOURNAL OF VIROLOGY., vol. 68, December 1994 (1994-12-01), pages 8254 - 8264, XP002094213 *
SMITH, C. ET AL.: "Transient protection of human T-cells from human immunodeficiency virus type 1 infection by transduction with adeno-associated viral vectors which express RNA decoys", ANTIVIRAL RESEARCH, vol. 32, 1996, pages 99 - 115, XP002094211 *
SYMENSMA, T. ET AL.: "RNA aptamers selected to bind human immunodeficiency virus type 1 rev in vitro are rev responsive in vivo", JOURNAL OF VIROLOGY., vol. 70, January 1996 (1996-01-01), pages 179 - 187, XP002094212 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002101036A1 (fr) * 2001-05-11 2002-12-19 Kazunari Taira Methode de formation d'un complexe stable d'un transcrit d'adn codant un peptide arbitraire a l'aide d'un produit de traduction, construction d'acide nucleique utilisee dans cette methode, complexe obtenu par cette methode et criblage de proteine fonctionnelle et marn ou adn codant la proteine a l'aide de cette methode
WO2016142948A1 (fr) 2015-03-11 2016-09-15 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Oligonucléotides leurres pour le traitement de maladies
US10781445B2 (en) 2015-03-11 2020-09-22 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Decoy oligonucleotides for the treatment of diseases

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