WO2003093424A3 - In vivo ssdna expression vectors for altering gene expression - Google Patents

In vivo ssdna expression vectors for altering gene expression Download PDF

Info

Publication number
WO2003093424A3
WO2003093424A3 PCT/US2003/013593 US0313593W WO03093424A3 WO 2003093424 A3 WO2003093424 A3 WO 2003093424A3 US 0313593 W US0313593 W US 0313593W WO 03093424 A3 WO03093424 A3 WO 03093424A3
Authority
WO
WIPO (PCT)
Prior art keywords
ssdna
stem
stranded
host cell
sequence
Prior art date
Application number
PCT/US2003/013593
Other languages
French (fr)
Other versions
WO2003093424A2 (en
WO2003093424A9 (en
Inventor
Yin Chen
Charles A Conrad
Original Assignee
Cytogenix Inc
Yin Chen
Charles A Conrad
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
Priority claimed from US10/136,218 external-priority patent/US20030082800A1/en
Application filed by Cytogenix Inc, Yin Chen, Charles A Conrad filed Critical Cytogenix Inc
Priority to AU2003265907A priority Critical patent/AU2003265907A1/en
Priority to US10/513,191 priority patent/US20050260588A1/en
Publication of WO2003093424A2 publication Critical patent/WO2003093424A2/en
Publication of WO2003093424A3 publication Critical patent/WO2003093424A3/en
Publication of WO2003093424A9 publication Critical patent/WO2003093424A9/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2121/00—Preparations for use in 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
    • C12N2840/00—Vectors comprising a special translation-regulating system
    • C12N2840/20—Vectors comprising a special translation-regulating system translation of more than one cistron

Landscapes

  • Genetics & Genomics (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

An expression vector for altering expression of a target nucleic acid sequence in a host cell by production of single-stranded cDNA (ssDNA) in the host cell in vivo. The expression vector is comprised of a cassette comprising a sequence of interest, an inverted tandem repeat, and a primer binding site 3' to the inverted tandem repeat, and a reverse transcriptase/RNAse H coding gene, and may be transfected into the host cell. Transcription of the cassette by the host cell produces an RNA template which is reverse transcribed with the product of the RT coding gene to produce ssDNA of a specified sequence. The ssDNA is modified to remove flanking vector sequences by taking advantage of the 'stem-loop' structure of the ssDNA, which forms as a result of the inverted tandem repeat that allows the ssDNA to fold back on itself, forming a double stranded DNA stem. The double-stranded stem may contain one or more restriction endonuclease recognition sites and the loop, which remains as ssDNA, can be any desired nucleotide sequence. This design allows the double-stranded stem of the stem-loop intermediate to be cleaved by the desired corresponding restriction endonuclease(s) and the loop portion is then released as a linearized, single-stranded piece of DNA. The resulting ssDNA binds to an endogenous target nucleic acid sequence to alter the expression of that sequence for such therapeutic purposes as gene activation or inactivation using duplex or triplex binding of nucleic acids, site-directed mutagenesis, interruption of cellular function by binding to specific cellular proteins, or interfering with RNA splicing functions.
PCT/US2003/013593 2002-05-01 2003-05-01 In vivo ssdna expression vectors for altering gene expression WO2003093424A2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU2003265907A AU2003265907A1 (en) 2002-05-01 2003-05-01 In vivo ssdna expression vectors for altering gene expression
US10/513,191 US20050260588A1 (en) 2002-05-01 2003-05-01 In vivo ssdna expression vectors for altering gene expression

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US10/136,218 2002-05-01
US10/136,218 US20030082800A1 (en) 1998-10-09 2002-05-01 In vivo ssDNA expression vectors for altering gene expression
US13621803A 2003-05-01 2003-05-01

Publications (3)

Publication Number Publication Date
WO2003093424A2 WO2003093424A2 (en) 2003-11-13
WO2003093424A3 true WO2003093424A3 (en) 2004-03-25
WO2003093424A9 WO2003093424A9 (en) 2004-06-10

Family

ID=31996565

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2003/013593 WO2003093424A2 (en) 2002-05-01 2003-05-01 In vivo ssdna expression vectors for altering gene expression

Country Status (1)

Country Link
WO (1) WO2003093424A2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7419964B2 (en) 1999-09-16 2008-09-02 Cytogenix, Inc. Treatment of HSV-related pathologies using ssDNA

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0532380A2 (en) * 1991-08-30 1993-03-17 University Of Medicine And Dentistry Of New Jersey Method for synthesizing stable single-stranded cDNA in eukaryotes by means of bacterial retron, products and uses therefor
EP0562206A2 (en) * 1992-01-06 1993-09-29 University Of Medicine & Dentistry Of New Jersey Single-stranded DNA-RNA hybrid molecules and reverse transcriptases
WO2000022114A1 (en) * 1998-10-09 2000-04-20 Ingene, Inc. PRODUCTION OF ssDNA $i(IN VIVO)
US20030082800A1 (en) * 1998-10-09 2003-05-01 Cytogenix, Inc. In vivo ssDNA expression vectors for altering gene expression

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5436141A (en) * 1989-02-24 1995-07-25 University Of Medicine And Dentistry Of New Jersey Method for synthesizing stable single-stranded CDNA in eukaryotes by means of a bacterial retron and products
EP0532380A2 (en) * 1991-08-30 1993-03-17 University Of Medicine And Dentistry Of New Jersey Method for synthesizing stable single-stranded cDNA in eukaryotes by means of bacterial retron, products and uses therefor
EP0562206A2 (en) * 1992-01-06 1993-09-29 University Of Medicine & Dentistry Of New Jersey Single-stranded DNA-RNA hybrid molecules and reverse transcriptases
WO2000022114A1 (en) * 1998-10-09 2000-04-20 Ingene, Inc. PRODUCTION OF ssDNA $i(IN VIVO)
US20030082800A1 (en) * 1998-10-09 2003-05-01 Cytogenix, Inc. In vivo ssDNA expression vectors for altering gene expression

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
MOROCHNITCHENKO O. ET AL.: "Production of single-stranded DNA in mammalian cells by means of a bacterial retron", THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 269, no. 4, 28 January 1994 (1994-01-28), pages 2380 - 2383, XP002132577 *

Also Published As

Publication number Publication date
WO2003093424A2 (en) 2003-11-13
WO2003093424A9 (en) 2004-06-10

Similar Documents

Publication Publication Date Title
WO2006012221A3 (en) Target cell-specific short interfering rna and methods of use thereof
WO2002014472A3 (en) Improved methods of gene silencing using inverted repeat sequences
WO2004058940A3 (en) Sirna-mediated gene silencing
WO2003012054A3 (en) Methods and compositions relating to improved lentiviral vector production systems
WO2005049846A3 (en) Mutant herpes simplex virus and use thereof in the treatment of squamous cell cancer
WO2004065613A3 (en) Inducible small interfering rna (sirna) expression constructs for targeted gene silencing
WO2004007684A3 (en) Synthetic tag genes
CA2647264A1 (en) Pathogen-inducible synthetic promoter for plant expression
CA2553976A1 (en) Method for producing minus-strand rna viral vectors using hybrid promoter comprising cytomegalovirus enhancer and chicken .beta.-actin promoter
JP2005501557A5 (en)
WO2002000894A3 (en) Gene silencing vector
WO2004029212A3 (en) In vivo gene silencing by chemically modified and stable sirna
Beaudry et al. An efficient strategy for the synthesis of circular RNA molecules.
WO2006013103A3 (en) Inducible gene expression
WO2003008575A3 (en) Dna sequences for regulating transcription
WO2000060115A3 (en) Method for identifying accessible binding sites on rna
ID26107A (en) METHODS FOR AMPLIFICATION NOT SPECIFIC OF NUCLEIC ACID
MXPA02003422A (en) ALTERING GENE EXPRESSION WITH ssDNA PRODUCED IN VIVO.
WO2019140116A3 (en) Amplifiable rnas for therapeutic cell systems
WO2003104470A3 (en) Retrons for gene targeting
WO2002018586A1 (en) Design and construction of transcription template for synthesizing cell-free protein, and dilution batch-type method of synthesizing cell-free wheat germ protein by using the same
IL145588A0 (en) Amplification and sequencing primer pairs and use thereof
AU2022234607A1 (en) Self-circularized RNA structure
WO2003004703A1 (en) Process for producing template dna and process for producing protein in cell-free protein synthesis system with the use of the same
US11713460B2 (en) Protecting RNAs from degradation using engineered viral RNAs

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ OM PH PL PT RO RU SD SE SG SK SL TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
COP Corrected version of pamphlet

Free format text: PAGES 1-31, DESCRIPTION, REPLACED BY NEW PAGES 1-46; PAGE 32, CLAIMS, REPLACED BY A NEW PAGE 47

122 Ep: pct application non-entry in european phase
WWE Wipo information: entry into national phase

Ref document number: 10513191

Country of ref document: US

NENP Non-entry into the national phase

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP