Methods and compositions for reducing screening in oligonucleotide-directed nucleic acid sequence alteration
OwnerEric B. Kmiec, Anja van Brabantinventors
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Abstract
Presented are methods, compositions and kits for reducing the number of target nucleic acid molecules required to be screened during oligonucleotide-directed nucleic acid sequence alteration.
Claims20
A method for identifying a cell having a desired oligonucleotide-directed sequence alteration at a first nucleic acid target site within the cell, the method comprising:
- identifying said desired sequence alteration in cells that have been selected for the presence of a selectable phenotype conferred by a concurrent oligonucleotide-directed sequence alteration at a second nucleic acid target site within said cells.
A method for effecting a desired sequence alteration at a first nucleic acid target site within a cell, the method comprising:
- concurrently targeting first and second nucleic acid sites within said cell for sequence alteration with respective first and second sequence-altering oligonucleotides, wherein said second alteration confers a selectable phenotype upon said cell;
- selecting cells having said selectable phenotype;
- and then
- identifying among the selected cells a cell having the desired sequence alteration at said first nucleic acid target site.
A composition for effecting a desired sequence alteration at a first nucleic acid target site within a cell, comprising:
- first and second sequence-altering oligonucleotides, wherein the oligonucleotides are capable of effecting sequence alteration at the first nucleic acid target site and at a second nucleic acid target site, respectively;
- and
- wherein alteration of the second nucleic acid target site confers a selectable phenotype.
A kit for effecting a desired sequence alteration at a first nucleic acid target site within a cell, comprising:
- first and second sequence-altering oligonucleotides, wherein the oligonucleotides are capable of effecting sequence alteration at the first nucleic acid target site and at a second nucleic acid target site, respectively;
- and
- wherein alteration of the second nucleic acid target site confers a selectable phenotype.
Dependencies are inferred from the claim text.