Mammalian artificial chromosomes
OwnerTsuneko Okazaki, Hiroshi Masumoto +3inventors
InventorsTsuneko Okazaki, Hiroshi Masumoto, Masashi Ikeno, Howard J. Cooke, Brenda Rose Grimes
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Abstract
A mammalian chromosome is produced according to the method comprising the steps of: introducing a DNA construct comprising a mammalian telomere and a centromere into a mammalian cell, wherein the centromere has a DNA sequence containing a plurality of copies of the CENP-B box sequence consisting of: 5′-NTTCGNNNNANNCGGGN-3′, wherein N is any one of A, T, C and G.
Claims54
A DNA construct comprising:
- a mammalian telomere and
- a DNA sequence of a mammalian centromere,
- wherein the DNA construct forms an artificial chromosome when introduced into mammalian cells, autonomously replicates in mammalian cells, is maintained extra chromosomally and is transmitted to progeny cells thereof, and wherein said DNA sequence contains a plurality of copies of the sequence:
- 5′-NTTCGNNNNANNCGGGN-3′,
- wherein N is selected from the group consisting of A, T, C and G.
A recombinant DNA construct comprising:
- a mammalian telomere,
- a mammalian centromere comprising tandem repeats of the sequence:
- 5′-NTTCGNNNNANNCGGGN-3′,
- wherein N is any one of A,T,C and G and wherein the sequence is provided in a quantity that causes the DNA construct to form a mammalian artificial chromosome when introduced into a mammalian cell.
A recombinant DNA construct comprising:
- a mammalian telomere,
- a mammalian centromere comprising tandem repeats of the sequence designated as Seq. No. 2, and wherein the sequence is provided in a quantity that causes the DNA construct to form a mammalian artificial chromosome when introduced into a mammalian cell.
A recombinant DNA construct comprising:
- a mammalian telomere,
- mammalian centromere comprising a sequence that is capable of hybridizing under selective conditions with a sequence selected from the group consisting of Seq. No. 2, Seq. No. 3, Seq. No. 4, Seq. No. 5, Seq. No. 6 and Seq. No. 7.
A cell line deposited at the Agency of Industrial Science and Technology, National Institute of Bioscience and Human Technology and designated as FERM BP-5625.
A method of performing homologous recombination comprising:
- transfecting a recombinant DNA construct into a host cell that does not possess the gene for DNA recombination within its DNA, wherein the recombinant DNA construct may be carried during mitotic division of the host cell within a plasmid having a gene for DNA recombination, which plasmid is capable of being eliminated during mitotic division of the host cell, and
- collecting host cells that possess the recombinant DNA construct, but do not possess the plasmid.
Dependencies are inferred from the claim text.