Yeast artificial chromosomes and their use in the control of gene expression

• US 6,348,349 B1
• Filed: 07/06/1998
• Issued: 02/19/2002
• Est. Priority Date: 09/10/1991
• Status: Expired due to Term

• Current Assignee: Crescendo Biologics Limited
• Original Assignee: The Babraham Institute
• Inventors: Bruggemann, Marianne
• Primary Examiner(s): Brusca, John S.
• Application Number:
  US09/110,811
• Time in Patent Office: 1,324 Days
• Field of Search: 435/320.1, 435/325, 530/23.1, 530/23.2, 530/23.7
• US Class Current: 435/325
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

Embryonic stem cells or other cells (not prokaryotic or yeast) that are essentially free of yeast DNA are prepared from suitably marked yeast artificial chromosomes and used to transfer DNA segments of considerable size into organisms.

Claims

1. A murine embryonic stem cell, transformed with a foreign gene or gene locus of at least 100 kb wherein said cell is essentially free of yeast DNA.

2. The cell according to claim 1, wherein said cell further comprises a marker gene.

3. The cell according to claim 2, comprising at least two copies of said marker gene.

4. The cell according to claim 2, wherein said marker is for resistance to neomycin, hygromycin, or HPRT.

5. The cell according to claim 2, wherein the marker gene is in addition to an ampicillin-resistance or other prokaryotic selective marker gene and/or uracil-resistance or other yeast-selective marker gene.

6. The cell according to claim 1, wherein the foreign gene or gene locus is in germ line configuration.

7. The cell according to claim 1, wherein the foreign gene or gene locus is of al least 1 Mb.

8. A murine embryonic stem cell, comprising a foreign gene or gene locus of at least 100 kb and a marker gene, separate from the foreign gene or gene locus, which allows selection by one or more properties in the cell, wherein said cell is essentially free of yeast DNA.