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DE12716101T1 - Adam6-mäuse - Google Patents

Adam6-mäuse Download PDF

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Publication number
DE12716101T1
DE12716101T1 DE12716101T DE12716101T DE12716101T1 DE 12716101 T1 DE12716101 T1 DE 12716101T1 DE 12716101 T DE12716101 T DE 12716101T DE 12716101 T DE12716101 T DE 12716101T DE 12716101 T1 DE12716101 T1 DE 12716101T1
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mouse
heavy chain
immunoglobulin
locus
human
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Sean Stevens
Andrew J. Murphy
Lynn MacDonald
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Regeneron Pharmaceuticals Inc
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Regeneron Pharmaceuticals Inc
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Abstract

Eine Maus, die ein Genom hat, das eine Veränderung eines Locus einer schweren Kette eines Immunglobulins umfasst, wobei die Veränderung endogene ADAM6-Funktion reduziert oder eliminiert, und die Maus weiter eine Nukleinsäuresequenz umfasst, die ein Maus-ADAM6-Protein oder ein Ortholog oder Homolog davon kodiert oder ein funktionelles Fragment des korrespondierenden ADAM6-Proteins.

Claims (30)

  1. Eine Maus, die ein Genom hat, das eine Veränderung eines Locus einer schweren Kette eines Immunglobulins umfasst, wobei die Veränderung endogene ADAM6-Funktion reduziert oder eliminiert, und die Maus weiter eine Nukleinsäuresequenz umfasst, die ein Maus-ADAM6-Protein oder ein Ortholog oder Homolog davon kodiert oder ein funktionelles Fragment des korrespondierenden ADAM6-Proteins.
  2. Die Maus von Anspruch 1, wobei die Nukleinsäuresequenz an einer ektopischen Position positioniert ist.
  3. Die Maus von Anspruch 1 oder 2, wobei die Nukleinsäuresequenz an einem endogenen Immunglobulin-Locus ist.
  4. Die Maus von Anspruch 1 oder 2, wobei die Nukleinsäuresequenz an einer anderen Position als einem endogenen Immunglobulin-Locus in das Mausgenom integriert ist.
  5. Die Maus irgendeines der Ansprüche 1–4, wobei die Veränderung des Locus der schweren Immunglobulin-Kette das Platzieren einer oder mehrerer humaner Immunglobulin-Gensequenzen umfasst.
  6. Die Maus irgendeines der Ansprüche 1–4, wobei die Veränderung des Locus der schweren Immunglobulin-Kette das Ersetzen einer oder mehrerer Sequenzen in einem Locus einer schweren Maus-Immunglobulin-Kette durch eine oder mehrere humane Immunglobulin-Gensequenzen umfasst.
  7. Die Maus von Anspruch 6, wobei die Veränderung des Lokus der schweren Immunglobulin-Kette das Ersetzen eines oder mehrerer endogener schwere Kette V (VH)-Gensegmente durch ein oder mehrere humane schwere Kette V (VH)-Gensegmente umfasst.
  8. Die Maus von Anspruch 1, wobei die Veränderung des Lokus der schweren Immunglobulin-Kette das Ersetzen einer endogenen schwere Kette variablen Gensequenz durch eine humane schwere Kette variablen Gensequenz umfasst.
  9. Die Maus irgendeines der Ansprüche 1–8, wobei die Nukleinsäuresequenz ein Maus-ADAM6a-Protein und/oder ADAM6b-Protein oder ein Ortholog, ein Homolog oder ein funktionelles Fragment davon kodiert.
  10. Ein Verfahren zur Veränderung eines Lokus der schweren Immunglobulin-Kette einer Maus, umfassend: (a) Vornehmen einer ersten Veränderung des Lokus der schweren Maus-Immunglobulin-Kette, die zu einer Reduktion oder Elimination endogener Maus-ADAM6 Aktivität in einer männlichen Maus führt, und (b) Vornehmen einer zweiten Veränderung der Maus, um eine Nukleinsäuresequenz hinzuzufügen, die der Maus ADAM6 Aktivität verleiht, die in einer männlichen Maus funktional ist.
  11. Das Verfahren gemäß Anspruch 10, wobei die Nukleinsäuresequenz in Schritt (b) an einer ektopischen Position zugefügt wird.
  12. Das Verfahren gemäß Anspruch 10 oder 11, wobei die erste Veränderung das Platzieren einer oder mehrerer humaner Immunglobulin-Gensequenzen umfasst.
  13. Das Verfahren gemäß Anspruch 10 oder 11, wobei die erste Veränderung das Ersetzen einer oder mehrerer Sequenzen in dem Locus einer schweren Maus-Immunglobulin-Kette durch eine oder mehrere humane Immunglobulin-Gesequenzen umfasst.
  14. Das Verfahren gemäß Anspruch 13 wobei die erste Veränderung das Ersetzen eines oder mehrerer endogener VH-Gensegmente durch ein oder mehrere humane VH-Gensegmente umfasst.
  15. Das Verfahren gemäß Anspruch 10, wobei die erste Veränderung das Ersetzen einer endogenen schwere Kette variablen Gensequenz durch eine humane schwere Kette variable Gensequenz umfasst.
  16. Das Verfahren gemäß irgendeinem der Ansprüche 10–15, wobei die erste und zweite Veränderung gleichzeitig vorgenommen werden.
  17. Eine durch das Verfahren irgendeines der Ansprüche 10–16 erhältliche Maus.
  18. Eine isolierte Zelle aus einer Maus irgendeines der Ansprüche 1–9 und 17.
  19. Ein isoliertes Gewebe aus einer Maus irgendeines der Ansprüche 1–9 und 17.
  20. Verwendung einer Maus gemäß irgendeines der Ansprüche 1–9 und 17 zur Herstellung eines human-Maus revers chimären Antikörpers, eines vollhumanen Antikörpers, eines vollhumanen Fab-Fragments und/oder eines vollhumanen F(ab)2-Fragments.
  21. Ein Verfahren zum Herstellen eines revers chimären Maus-humanen Antikörpers, der spezifisch gegen ein Antigen ist, umfassend die Schritte: a) Immunisieren einer Maus gemäß irgendeinem der Ansprüche 1–9 und 17 mit dem Antigen; b) Isolieren wenigstens einer Zelle von der Maus, die einen revers chimären Maus-humanen Antikörper, der spezifisch gegen das Antigen ist, produziert; und c) Kultivieren wenigstens einer Antikörper-produzierenden Zelle des Schritts b) und Erhalten des Antikörpers.
  22. Das Verfahren von Anspruch 21, wobei das Kultivieren in Schritt c) an wenigstens einer Hybridomzelle, die aus der wenigstens einen in Schritt b) erhaltenen Zelle hergestellt ist, durchgeführt wird.
  23. Ein Verfahren zum Herstellen eines vollhumanen Antikörpers, der spezifisch gegen ein Antigen ist, umfassend die Schritte: a) Immunisieren einer Maus gemäß irgendeinem der Ansprüche 1–9 und 17 mit dem Antigen; b) Isolieren wenigstens einer Zelle von der Maus, die einen revers chimären Maus-humanen Antikörper, der spezifisch gegen das Antigen ist, produziert; c) Herstellen wenigstens einer Zelle, die einen vollhumanen Antikörper, der von dem Antikörper des Schritts b) abgeleitet und spezifisch gegen das Antigen ist; und d) Kultivieren wenigstens einer Antikörper-produzierenden Zelle des Schritts c) und Erhalten des Antikörpers.
  24. Das Verfahren irgendeines der Ansprüche 21 und 23, wobei die wenigstens eine in Schritt b) erhaltene Zelle ein Splenozyt oder eine B-Zelle ist.
  25. Das Verfahren irgendeines der Ansprüche 21–24, wobei der Antikörper ein monoklonaler Antikörper ist.
  26. Das Verfahren irgendeines der Ansprüche 21–25, wobei das Immunisieren mit dem Antigen des Schritts a) mit Protein, DNS, einer Kombination von DNS und Protein, oder mit das Antigen exprimierenden Zellen durchgeführt wird.
  27. Verwendung einer Nukleinsäuresequenz, die ein Maus-ADAM6-Protein, oder ein Ortholog oder ein Homolog davon kodiert, oder ein funktionelles Fragment des korrespondierenden ADAM6-Proteins, zum Wiederherstellen oder Verbessern der Fertilität einer Maus, die ein eine Veränderung eines Locus der schweren Immunglobulin-Kette umfassendes Genom hat, wobei die Veränderung die endogene ADAM6-Funktion reduziert oder eliminiert.
  28. Die Verwendung des Anspruchs 27, wobei die Nukleinsäuresequenz an einer ektopischen Position in das Genom der Maus integriert wird.
  29. Die Verwendung von Anspruch 27 oder 28, wobei die Nukleinsäuresequenz an einem endogenen Immunglobulin-Locus in das Genom der Maus integriert wird.
  30. Die Verwendung von Anspruch 27 oder 28, wobei die Nukleinsäuresequenz an einer anderen Position als einem endogenen Immunglobulin-Locus in das Genom der Maus integriert wird.
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