Crispr oligoncleotides and gene editing
AssigneesLife Technologies Corporation, Thermo Fisher Scientific Geneart GmbH
InventorsNamritha RAVINDER, Korbinian HEIL, Yizhu GUO, Xiquan LIANG, Robert POTTER, Sanjay KUMAR
Life of this patent
Estimating the term…
Abstract
The present disclosure generally relates to compositions and methods for the genetic modification of cells. In particular, the disclosure relates to CRISPR reagents and the use of such reagents.
Claims16
A method for producing a nucleic acid molecule, the method comprising performing polymerase chain reaction (PCR) in a reaction mixture containing (i) a double-stranded nucleic acid segment and (ii) at least one oligonucleotide capable of hybridizing to nucleic acid at one terminus of the double-stranded nucleic acid segment,
- wherein the nucleic acid molecule is produced by the PCR reaction, and
- wherein the product nucleic acid molecule contains at or near one terminus a promoter suitable for in vitro transcription.
-26. (canceled)
A ligated RNA molecule comprising two RNA regions that differ in nucleotide sequence connected by a ligation group, wherein the ligated RNA molecule is capable of binding to a Cas9 protein and has a region of sequence complementarity of at least 10 nucleotides to a target locus.
A method for performing homologous recombination in a cell, the method comprising:
- (a) introducing into the cell a nucleic acid cutting entity capable of generating a double-stranded break at a specified location in a nucleic acid molecule present inside a cell to produce a cleaved nucleic acid molecule, and
- (b) introducing a donor nucleic acid molecule into the cell,
- wherein step (a) is performed before step (b) or wherein step (b) is performed before step (a).
Dependencies are inferred from the claim text.