DOI: 10.1073/pnas.96.3.1013. Generation of conditional mutants in higher eukaryotes by switching between the expression of two genes

Generation of conditional mutants in higher eukaryotes by switching between the expression of two genes

10.1073/pnas.96.3.1013

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

A regulatory system for the in-depth study of gene functions in higher eukaryotic cells has been developed. It is based on the tetracycline-controlled transactivators and reverse tTA, which were remodeled to discriminate efficiently between two different promoters. The system permits one to control reversibly the activity of two genes, or two alleles of a gene, in a mutually exclusive way, and also allows one to abrogate the activities of both. This dual regulatory circuit, which can be operated by a single effector substance such as doxycycline, overcomes limitations of conventional genetic approaches. The conditional mutants that can now be generated will be useful for the study of gene function in vitro and in vivo . In addition, the system may be of value for a variety of practical applications, including gene therapy.

Bibliography

Baron, U., Schnappinger, D., Helbl, V., Gossen, M., Hillen, W., & Bujard, H. (1999). Generation of conditional mutants in higher eukaryotes by switching between the expression of two genes. Proceedings of the National Academy of Sciences, 96(3), 1013â1018.

Authors 6

Udo Baron (first)
Dirk Schnappinger (additional)
Vera Helbl (additional)
Manfred Gossen (additional)
Wolfgang Hillen (additional)
Hermann Bujard (additional)

References 29 Referenced 81

10.1073/pnas.89.12.5547
10.1126/science.7792603
10.1016/S0076-6879(97)83014-5
10.1007/s11626-998-0043-8
10.1002/(SICI)1097-0061(199707)13:9<837::AID-YEA145>3.0.CO;2-T
10.1093/emboj/17.7.2079
10.1046/j.1365-313X.1994.5040559.x
10.1007/BF00017815
10.1073/pnas.93.20.10933
10.1073/pnas.92.8.3576
10.1126/science.273.5280.1384
10.1126/science.274.5293.1678
Kolster K. Kistner A. Baron U. Freundlieb S. Gossen M. & Bujard H. (1998) Transgenic Techniques in press.
10.1172/JCI117174
10.1093/nar/25.14.2723
10.1128/mcb.14.3.1669-1679.1994
P J Southern, P Berg J Mol Appl Genet 1, 327–341 (1982). / J Mol Appl Genet by Southern P J (1982)
E Harlow, D Lane Antibodies: A Laboratory Manual (Cold Spring Harbor Lab. Press, Plainview, NY, 1988). / Antibodies: A Laboratory Manual by Harlow E (1988)
10.1016/0022-2836(92)91065-W
10.1006/jmbi.1994.0044
10.1006/jmbi.1997.1539
10.1006/jmbi.1997.1540
10.1016/0378-1119(94)90130-9
10.1146/annurev.mi.48.100194.002021
10.1126/science.8153629
10.1002/j.1460-2075.1988.tb03290.x
U Baron Ph.D. thesis (Univ. Heidelberg, 1998). / Ph.D. thesis by Baron U (1998)
10.1073/pnas.95.23.13670
Freundlieb S. Schirra Ch. & Bujard H. (1999) J. Gene Medicine in press.

Dates

Type	When
Created	23 years ago (July 26, 2002, 10:40 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 5:27 p.m.)
Indexed	1 month, 3 weeks ago (June 30, 2025, 7:26 a.m.)
Issued	26 years, 6 months ago (Feb. 2, 1999)
Published	26 years, 6 months ago (Feb. 2, 1999)
Published Online	26 years, 6 months ago (Feb. 2, 1999)
Published Print	26 years, 6 months ago (Feb. 2, 1999)

Funders 0

None

BibTeX

@article{Baron_1999, title={Generation of conditional mutants in higher eukaryotes by switching between the expression of two genes}, volume={96}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.96.3.1013}, DOI={10.1073/pnas.96.3.1013}, number={3}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Baron, Udo and Schnappinger, Dirk and Helbl, Vera and Gossen, Manfred and Hillen, Wolfgang and Bujard, Hermann}, year={1999}, month=feb, pages={1013–1018} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        6,
        30
      ]
    ],
    "date-time": "2025-06-30T11:26:38Z",
    "timestamp": 1751282798888
  },
  "reference-count": 29,
  "publisher": "Proceedings of the National Academy of Sciences",
  "issue": "3",
  "content-domain": {
    "domain": [
      "www.pnas.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        1999,
        2,
        2
      ]
    ]
  },
  "abstract": "<jats:p>\n            A regulatory system for the in-depth study of gene functions in higher eukaryotic cells has been developed. It is based on the tetracycline-controlled transactivators and reverse tTA, which were remodeled to discriminate efficiently between two different promoters. The system permits one to control reversibly the activity of two genes, or two alleles of a gene, in a mutually exclusive way, and also allows one to abrogate the activities of both. This dual regulatory circuit, which can be operated by a single effector substance such as doxycycline, overcomes limitations of conventional genetic approaches. The conditional mutants that can now be generated will be useful for the study of gene function\n            <jats:italic>in vitro</jats:italic>\n            and\n            <jats:italic>in vivo</jats:italic>\n            . In addition, the system may be of value for a variety of practical applications, including gene therapy.\n          </jats:p>",
  "DOI": "10.1073/pnas.96.3.1013",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        26
      ]
    ],
    "date-time": "2002-07-26T14:40:52Z",
    "timestamp": 1027694452000
  },
  "page": "1013-1018",
  "update-policy": "http://dx.doi.org/10.1073/pnas.cm10313",
  "source": "Crossref",
  "is-referenced-by-count": 81,
  "title": "Generation of conditional mutants in higher eukaryotes by switching between the expression of two genes",
  "prefix": "10.1073",
  "volume": "96",
  "author": [
    {
      "given": "Udo",
      "family": "Baron",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Zentrum f\u00fcr Molekulare Biologie, Universit\u00e4t Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Lehrstuhl f\u00fcr Mikrobiologie, Universit\u00e4t Erlangen, Staudtstrasse 5, D-91058 Erlangen, Germany; and University of California, MCB Barker/Koshland ASU, 401 Barker Hall, Berkeley, CA 94720-3202"
        }
      ]
    },
    {
      "given": "Dirk",
      "family": "Schnappinger",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Zentrum f\u00fcr Molekulare Biologie, Universit\u00e4t Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Lehrstuhl f\u00fcr Mikrobiologie, Universit\u00e4t Erlangen, Staudtstrasse 5, D-91058 Erlangen, Germany; and University of California, MCB Barker/Koshland ASU, 401 Barker Hall, Berkeley, CA 94720-3202"
        }
      ]
    },
    {
      "given": "Vera",
      "family": "Helbl",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Zentrum f\u00fcr Molekulare Biologie, Universit\u00e4t Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Lehrstuhl f\u00fcr Mikrobiologie, Universit\u00e4t Erlangen, Staudtstrasse 5, D-91058 Erlangen, Germany; and University of California, MCB Barker/Koshland ASU, 401 Barker Hall, Berkeley, CA 94720-3202"
        }
      ]
    },
    {
      "given": "Manfred",
      "family": "Gossen",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Zentrum f\u00fcr Molekulare Biologie, Universit\u00e4t Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Lehrstuhl f\u00fcr Mikrobiologie, Universit\u00e4t Erlangen, Staudtstrasse 5, D-91058 Erlangen, Germany; and University of California, MCB Barker/Koshland ASU, 401 Barker Hall, Berkeley, CA 94720-3202"
        }
      ]
    },
    {
      "given": "Wolfgang",
      "family": "Hillen",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Zentrum f\u00fcr Molekulare Biologie, Universit\u00e4t Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Lehrstuhl f\u00fcr Mikrobiologie, Universit\u00e4t Erlangen, Staudtstrasse 5, D-91058 Erlangen, Germany; and University of California, MCB Barker/Koshland ASU, 401 Barker Hall, Berkeley, CA 94720-3202"
        }
      ]
    },
    {
      "given": "Hermann",
      "family": "Bujard",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Zentrum f\u00fcr Molekulare Biologie, Universit\u00e4t Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany; Lehrstuhl f\u00fcr Mikrobiologie, Universit\u00e4t Erlangen, Staudtstrasse 5, D-91058 Erlangen, Germany; and University of California, MCB Barker/Koshland ASU, 401 Barker Hall, Berkeley, CA 94720-3202"
        }
      ]
    }
  ],
  "member": "341",
  "published-online": {
    "date-parts": [
      [
        1999,
        2,
        2
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_3_3_1_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.89.12.5547"
    },
    {
      "key": "e_1_3_3_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.7792603"
    },
    {
      "key": "e_1_3_3_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0076-6879(97)83014-5"
    },
    {
      "key": "e_1_3_3_4_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/s11626-998-0043-8"
    },
    {
      "key": "e_1_3_3_5_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/(SICI)1097-0061(199707)13:9<837::AID-YEA145>3.0.CO;2-T"
    },
    {
      "key": "e_1_3_3_6_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/17.7.2079"
    },
    {
      "key": "e_1_3_3_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-313X.1994.5040559.x"
    },
    {
      "key": "e_1_3_3_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/BF00017815"
    },
    {
      "key": "e_1_3_3_9_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.93.20.10933"
    },
    {
      "key": "e_1_3_3_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.92.8.3576"
    },
    {
      "key": "e_1_3_3_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.273.5280.1384"
    },
    {
      "key": "e_1_3_3_12_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.274.5293.1678"
    },
    {
      "key": "e_1_3_3_13_2",
      "unstructured": "Kolster K. Kistner A. Baron U. Freundlieb S. Gossen M. & Bujard H. (1998) Transgenic Techniques in press."
    },
    {
      "key": "e_1_3_3_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1172/JCI117174"
    },
    {
      "key": "e_1_3_3_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/25.14.2723"
    },
    {
      "key": "e_1_3_3_16_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/mcb.14.3.1669-1679.1994"
    },
    {
      "key": "e_1_3_3_17_2",
      "first-page": "327",
      "volume": "1",
      "author": "Southern P J",
      "year": "1982",
      "unstructured": "P J Southern, P Berg J Mol Appl Genet 1, 327\u2013341 (1982).",
      "journal-title": "J Mol Appl Genet"
    },
    {
      "key": "e_1_3_3_18_2",
      "volume-title": "Antibodies: A Laboratory Manual",
      "author": "Harlow E",
      "year": "1988",
      "unstructured": "E Harlow, D Lane Antibodies: A Laboratory Manual (Cold Spring Harbor Lab. Press, Plainview, NY, 1988)."
    },
    {
      "key": "e_1_3_3_19_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0022-2836(92)91065-W"
    },
    {
      "key": "e_1_3_3_20_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1994.0044"
    },
    {
      "key": "e_1_3_3_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1997.1539"
    },
    {
      "key": "e_1_3_3_22_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1997.1540"
    },
    {
      "key": "e_1_3_3_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0378-1119(94)90130-9"
    },
    {
      "key": "e_1_3_3_24_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.mi.48.100194.002021"
    },
    {
      "key": "e_1_3_3_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.8153629"
    },
    {
      "key": "e_1_3_3_26_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/j.1460-2075.1988.tb03290.x"
    },
    {
      "key": "e_1_3_3_27_2",
      "volume-title": "Ph.D. thesis",
      "author": "Baron U",
      "year": "1998",
      "unstructured": "U Baron Ph.D. thesis (Univ. Heidelberg, 1998)."
    },
    {
      "key": "e_1_3_3_28_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.95.23.13670"
    },
    {
      "key": "e_1_3_3_29_2",
      "unstructured": "Freundlieb S. Schirra Ch. & Bujard H. (1999) J. Gene Medicine in press."
    }
  ],
  "container-title": "Proceedings of the National Academy of Sciences",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://pnas.org/doi/pdf/10.1073/pnas.96.3.1013",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2022,
        4,
        13
      ]
    ],
    "date-time": "2022-04-13T21:27:53Z",
    "timestamp": 1649885273000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://pnas.org/doi/full/10.1073/pnas.96.3.1013"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1999,
        2,
        2
      ]
    ]
  },
  "references-count": 29,
  "journal-issue": {
    "issue": "3",
    "published-print": {
      "date-parts": [
        [
          1999,
          2,
          2
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1073/pnas.96.3.1013"
  ],
  "URL": "http://dx.doi.org/10.1073/pnas.96.3.1013",
  "relation": {},
  "ISSN": [
    "0027-8424",
    "1091-6490"
  ],
  "subject": [],
  "container-title-short": "Proc. Natl. Acad. Sci. U.S.A.",
  "published": {
    "date-parts": [
      [
        1999,
        2,
        2
      ]
    ]
  },
  "assertion": [
    {
      "value": "1998-09-28",
      "order": 0,
      "name": "received",
      "label": "Received",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    },
    {
      "value": "1999-02-02",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}