Methods and compositions for the generation of humanized mice
First Claim
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1. A method of generating a humanized animal, comprising:
- recombining a first DNA construct with a second DNA construct, wherein the first DNA construct has a non-human animal DNA sequence contained therein, and wherein the second DNA construct has a human DNA sequence contained therein and the human DNA sequence is flanked by a first and a second non-human animal DNA sequence;
isolating a recombined third DNA construct having a human DNA sequence flanked by the first and second non-human animal DNA sequence; and
introducing the recombined third DNA construct into a non-human embryogenic stem cell.
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Abstract
The invention provides methods and compositions for generating non-human transgenic animals that are humanized at one or more gene sequences. According to the methods of the invention, a DNA construct containing a human DNA sequence flanked by sequences from the non-human animal is generated by recombination in a bacterial cell, for example, in E. coli. The DNA construct that is produced can then be introduced into a non-human embryogenic stem cell where it can recombine with the genomic DNA of the non-human animal.
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Citations
42 Claims
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1. A method of generating a humanized animal, comprising:
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recombining a first DNA construct with a second DNA construct, wherein the first DNA construct has a non-human animal DNA sequence contained therein, and wherein the second DNA construct has a human DNA sequence contained therein and the human DNA sequence is flanked by a first and a second non-human animal DNA sequence;
isolating a recombined third DNA construct having a human DNA sequence flanked by the first and second non-human animal DNA sequence; and
introducing the recombined third DNA construct into a non-human embryogenic stem cell. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 41, 42)
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18. A DNA construct for performing homologous recombination within a cell, the construct comprising:
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a human DNA coding sequence having at least one intron disposed therein;
a selection marker gene contained within said at least one intron;
a first and second non-human animal DNA sequences flanking the human DNA, wherein the non-human animal flanking sequences are homologous to sequences in the genome of the non-human animal that flank a gene orthologous to the human DNA coding sequence. - View Dependent Claims (19, 20, 21, 22, 23, 24)
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25. A method for generating a DNA construct for performing homologous recombination within a cell by
recombining a first DNA construct with a second DNA construct, wherein the first DNA construct has a non-human animal DNA sequence contained therein, wherein the second DNA construct has a human DNA sequence contained therein and the human DNA sequence is flanked by a first and a second non-human animal DNA sequence; -
isolating a recombined third DNA construct having a human DNA sequence flanked by the first and second non-human animal DNA sequence; and
introducing the recombined third DNA construct into a non-human embryogenic stem cell. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40)
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Specification