WO2001002553A3 - Dna binding protein and sequence as insulators having specific enhancer blocking activity for regulation of gene expression - Google Patents

Dna binding protein and sequence as insulators having specific enhancer blocking activity for regulation of gene expression Download PDF

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Publication number
WO2001002553A3
WO2001002553A3 PCT/US2000/010509 US0010509W WO0102553A3 WO 2001002553 A3 WO2001002553 A3 WO 2001002553A3 US 0010509 W US0010509 W US 0010509W WO 0102553 A3 WO0102553 A3 WO 0102553A3
Authority
WO
WIPO (PCT)
Prior art keywords
blocking activity
regulation
insulator
gene expression
binding protein
Prior art date
Application number
PCT/US2000/010509
Other languages
French (fr)
Other versions
WO2001002553A2 (en
Inventor
Adam C Bell
Adam G West
Gary Felsenfeld
Original Assignee
Us Health
Adam C Bell
Adam G West
Gary Felsenfeld
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 Us Health, Adam C Bell, Adam G West, Gary Felsenfeld filed Critical Us Health
Priority to CA002377929A priority Critical patent/CA2377929A1/en
Priority to JP2001508326A priority patent/JP2003504016A/en
Priority to AU46475/00A priority patent/AU776752B2/en
Priority to EP00928209A priority patent/EP1196612A2/en
Publication of WO2001002553A2 publication Critical patent/WO2001002553A2/en
Publication of WO2001002553A3 publication Critical patent/WO2001002553A3/en

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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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression

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  • Genetics & Genomics (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

An insulator element shown to be both necessary and sufficient for the enhancer blocking activity in human cells is described. The insulator element of the invention is a 42 base pair DNA molecule having specific enhancer blocking activity for regulation of gene expression. The insulator element has also been shown to be the core binding site for CTCF, a DNA binding protein that is highly conserved in vertebrates. In addition, an insulator element containing CTCF binding sites has been identified in the region of the Igf 2 locus which is methylated exclusively on the paternal allele. Methylation of the insulator sequence abolishes the ability of CTCF to bind to the insulator and results in loss of enhancer-blocking activity.
PCT/US2000/010509 1999-06-30 2000-04-19 Dna binding protein and sequence as insulators having specific enhancer blocking activity for regulation of gene expression WO2001002553A2 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
CA002377929A CA2377929A1 (en) 1999-06-30 2000-04-19 Dna binding protein and sequence as insulators having specific enhancer blocking activity for regulation of gene expression
JP2001508326A JP2003504016A (en) 1999-06-30 2000-04-19 DNA binding proteins and sequences as insulators with specific enhancer blocking activity for regulation of gene expression
AU46475/00A AU776752B2 (en) 1999-06-30 2000-04-19 DNA binding protein and sequence as insulators having specific enhancer blocking activity for regulation of gene expression
EP00928209A EP1196612A2 (en) 1999-06-30 2000-04-19 Dna binding protein and sequence as insulators having specific enhancer blocking activity for regulation of gene expression

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US14172899P 1999-06-30 1999-06-30
US60/141,728 1999-06-30

Publications (2)

Publication Number Publication Date
WO2001002553A2 WO2001002553A2 (en) 2001-01-11
WO2001002553A3 true WO2001002553A3 (en) 2001-05-03

Family

ID=22496966

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2000/010509 WO2001002553A2 (en) 1999-06-30 2000-04-19 Dna binding protein and sequence as insulators having specific enhancer blocking activity for regulation of gene expression

Country Status (5)

Country Link
EP (1) EP1196612A2 (en)
JP (1) JP2003504016A (en)
AU (1) AU776752B2 (en)
CA (1) CA2377929A1 (en)
WO (1) WO2001002553A2 (en)

Families Citing this family (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2003290453A1 (en) 2002-12-20 2004-07-14 Chromagenics B.V. Means and methods for producing a protein through chromatin openers that are capable of rendering chromatin more accessible to transcription factors
ES2354160T3 (en) * 2003-10-21 2011-03-10 Merck Serono Sa MINIMUM DNA SEQUENCE THAT ACTS AS A CHROMATIN INSULATION, AND ITS USE IN THE EXPRESSION OF PROTEINS.
US20060063231A1 (en) 2004-09-16 2006-03-23 Sangamo Biosciences, Inc. Compositions and methods for protein production
US20060195935A1 (en) 2004-11-08 2006-08-31 Chromagenics B.V. Selection of host cells expressing protein at high levels
EP1809750B1 (en) 2004-11-08 2012-03-21 ChromaGenics B.V. Selection of host cells expressing protein at high levels
US8039230B2 (en) 2004-11-08 2011-10-18 Chromagenics B.V. Selection of host cells expressing protein at high levels
US8999667B2 (en) 2004-11-08 2015-04-07 Chromagenics B.V. Selection of host cells expressing protein at high levels
US20060172382A1 (en) 2004-11-08 2006-08-03 Chromagenics B.V. Selection of host cells expressing protein at high levels
EP1996705B1 (en) 2006-03-20 2011-08-31 ChromaGenics B.V. Expression augmenting dna fragments, use thereof, and methods for finding thereof
WO2010046493A2 (en) * 2008-10-23 2010-04-29 Université de Lausanne Gene transfer vectors comprising at least one isolated dna molecule having insulator and or boundary properties and methods to identify the same
EP2414524B1 (en) 2009-04-03 2017-08-23 Centre National De La Recherche Scientifique Gene transfer vectors comprising genetic insulator elements and methods to identify genetic insulator elements
WO2015138852A1 (en) 2014-03-14 2015-09-17 University Of Washington Genomic insulator elements and uses thereof
US11279974B2 (en) 2014-12-01 2022-03-22 The Broad Institute, Inc. Method for in situ determination of nucleic acid proximity
MX2020003320A (en) 2017-10-10 2021-07-16 Procter & Gamble Sulfate free clear personal cleansing composition comprising low inorganic salt.
WO2020040046A1 (en) * 2018-08-21 2020-02-27 学校法人慶應義塾 Enhancer blocker and use thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996004390A1 (en) * 1994-07-29 1996-02-15 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Dna sequence which acts as a chromatin insulator element to protect expressed genes from cis-acting regulatory sequences in mammalian cells
WO1997011972A1 (en) * 1995-09-28 1997-04-03 The Trustees Of Columbia University In The City Of New York Chimeric dna-binding/dna methyltransferase nucleic acid and polypeptide and uses thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996004390A1 (en) * 1994-07-29 1996-02-15 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Dna sequence which acts as a chromatin insulator element to protect expressed genes from cis-acting regulatory sequences in mammalian cells
WO1997011972A1 (en) * 1995-09-28 1997-04-03 The Trustees Of Columbia University In The City Of New York Chimeric dna-binding/dna methyltransferase nucleic acid and polypeptide and uses thereof

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
BELL, A.C. & FELSENFELD, G.: "Methylation of a CTCF-dependent boundary controls imprinted expression of the Igf2 gene", NATURE, vol. 405, no. 6785, 25 May 2000 (2000-05-25), pages 482 - 485, XP002155514 *
BELL, A.C. ET AL.: "The Preotein CTCF Is Required for the Enhancer Blocking Activity of Vertebrate Insulators", CELL, vol. 98, no. 3, 6 August 1999 (1999-08-06), pages 387 - 396, XP000926124 *
CHUNG, J.H. ET AL.: "Characterization of the chicken beta-globin insulator", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA, vol. 94, no. 2, 21 January 1997 (1997-01-21), pages 575 - 580, XP002155512 *
DATABASE EMBL ROD 1 January 1900 (1900-01-01), STADNICK M. P., ET AL.: "RATTUS NORVEGICUS H19 GENE, 5' FLANKING REGION.", XP002155515, Database accession no. AF043428 *
FREVEL, M.A. ET AL.: "A potential imprint control element: identification of a conserved 42 bp sequence upstream of H19", TRENDS IN GENETICS, vol. 15, no. 6, June 1999 (1999-06-01), pages 216 - 218, XP004170206 *
ZUBAIR, M. ET AL.: "Structure and Expression of the Mouse L23mrp Gene Downstream of the Imprinted H19 Gene: Biallelic Expression and Lack of Interaction with the H19 Enhancers", GENOMICS, vol. 45, no. 2, 15 October 1997 (1997-10-15), pages 290 - 296, XP002155513 *

Also Published As

Publication number Publication date
CA2377929A1 (en) 2001-01-11
AU776752B2 (en) 2004-09-23
EP1196612A2 (en) 2002-04-17
WO2001002553A2 (en) 2001-01-11
AU4647500A (en) 2001-01-22
JP2003504016A (en) 2003-02-04

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