High efficiency gene targeting in mouse embryonic stem cells
First Claim
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1. A method for modifying a target DNA sequence in a mouse embryonic stem cell comprising:
- (a) introducing in vitro a targeting DNA sequence into the mouse embryonic stem cell derived from an inbred mouse strain, said targeting DNA sequence is isolated from said inbred mouse strain; and
(b) isolating in vitro the mouse embryonic stem cell whose target DNA sequence has been modified by incorporation of said targeting DNA sequence into a nonselectable gene of the target sequence.
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Abstract
The present invention provides novel methods for modifying the genome of an animal cell which typically comprise the steps of: constructing a DNA molecule in which desired sequence modifications are contained in a segment of DNA (a “targeting DNA”) that is substantially isogenic with a DNA in the cell genome (a “target DNA”); introducing the targeting DNA construct into the cell (e.g., by microinjection, electroporation, transfection, or calcium phosphate precipitation); and selecting cells in which the desired sequence modifications have been introduced into the genome via homologous recombination.
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23 Claims
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1. A method for modifying a target DNA sequence in a mouse embryonic stem cell comprising:
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(a) introducing in vitro a targeting DNA sequence into the mouse embryonic stem cell derived from an inbred mouse strain, said targeting DNA sequence is isolated from said inbred mouse strain; and
(b) isolating in vitro the mouse embryonic stem cell whose target DNA sequence has been modified by incorporation of said targeting DNA sequence into a nonselectable gene of the target sequence. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23)
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Specification