Back to results
US 12,674,213 B2Granted
Compositions and methods for quantifying integration of recombinant vector nucleic acid
AssigneeJanssen Biotech, Inc.
InventorsRajitha Doddareddy, Hsing-Yin Liu, Levi Gray-Rupp, Tong-Yuan Yang
Life of this patent
Estimating the term…
Abstract
In certain aspects, the disclosure relates to methods of quantifying integration of a recombinant vector nucleic acid into a target cell's genome. The present disclosure also provides compositions and kits, including particular primers and probes, for performing the quantitation.
Claims46
A method for quantifying integration of a recombinant vector nucleic acid into a cellular genome, the method comprising:
- (a) providing a biological sample comprising a host cellular genome;
- (b) amplifying genomic DNA of the biological sample with a quantitative amplification technique using a primer pair comprising a first oligonucleotide primer and a second oligonucleotide primer, wherein at least one oligonucleotide primer of the primer pair specifically hybridizes to an integrated recombinant vector polynucleotide sequence;
- and
- (c) detecting and/or quantifying the genomic nucleic acid that was amplified through step (b);
- wherein the first oligonucleotide primer comprises the nucleic acid sequence of SEQ ID NO:
- 1, SEQ ID NO:
- 5, or SEQ ID NO:
- 14, and
- wherein the second oligonucleotide primer comprises the nucleic acid sequence of SEQ ID NO:
- 2, SEQ ID NO:
- 6, or SEQ ID NO:
- 15.
9 more elements
A method of quantifying integration of a lentiviral vector nucleic acid into a cellular genome, the method comprising:
- (a) providing a biological sample comprising a host cellular genome;
- (b) amplifying genomic DNA of the biological sample with a quantitative amplification technique using a primer pair comprising a first oligonucleotide primer and a second oligonucleotide primer, wherein at least one oligonucleotide primer of the primer pair specifically hybridizes to an integrated lentiviral vector polynucleotide sequence;
- and
- (c) quantifying the genomic nucleic acid that was amplified through step (b), wherein quantifying the lentiviral vector nucleic acid integrated in the host cellular genome comprises comparing the ratio of amplified lentiviral vector nucleic acid to a reference,
- wherein the oligonucleotide primer pair comprise the nucleic acid sequence of SEQ ID NO:
- 1 and the nucleic acid sequence of SEQ ID NO:2, respectively, or the nucleic acid sequence of SEQ ID NO:5 and the nucleic acid sequence of SEQ ID NO:6, respectively, or the nucleic acid sequence of SEQ ID NO:14 and the nucleic acid sequence of SEQ ID NO:15, respectively.
3 more elements
Dependencies are inferred from the claim text.