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US 8,945,862 B2Granted
Triple-fusion constructs and methods of monitoring human embryonic stem cells
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsJoseph Ching-Ming Wu, Feng Cao, Sanjiv Sam Gambhir
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Abstract
Embodiments of the present disclosure include triple-fusion human embryonic stem cells, methods of imaging triple-fusion human embryonic stem cells, triple-fusion polynucleotides, triple-fusion proteins, methods of monitoring the progression of human embryonic stem cells, methods of making isolated triple-fusion human embryonic stem cells, and the like.
Claims14
A method of monitoring human embryonic stem cells (hESCs) comprising:
- providing human embryonic stem cells;
- transducing the hESCs with a triple-fusion expression vector to produce triple-fusion hESCs, wherein the triple-fusion expression vector comprises a firefly luciferase reporter gene (Fluc) having a nucleotide sequence selected from the group consisting of:
- SEQ ID NO:
- 1 and SEQ ID NO:
- 3, a red fluorescent reporter gene (RFP) having a nucleotide sequence of SEQ ID NO:
- 6, and a truncated HSV1-tK positron emission tomography (PET) reporter gene (HSV1-tTK) having a nucleotide sequence selected from the group consisting of:
- SEQ ID NO:
- 7 and SEQ ID NO:
- 14;
- and
- detecting fluorescence, bioluminescence, a nuclear signal from a PET reporter, or a combination thereof, emitted from the triple-fusion hESCs and any progeny of the triple-fusion hESCs.
8 more elements
A method of monitoring human embryonic stem cells (hESCs) in a subject comprising:
- providing triple-fusion hESCs having a triple-fusion expression vector comprising a firefly luciferase reporter gene (Fluc) having a nucleotide sequence selected from the group consisting of:
- SEQ ID NO:
- 1 and SEQ ID NO:
- 3, a red fluorescent reporter gene (RFP) having a nucleotide sequence of SEQ ID NO:
- 6, and a truncated HSV1-tK positron emission tomography (PET) reporter gene (HSV1-tTK) having a nucleotide sequence selected from the group consisting of:
- SEQ ID NO:
- 7 and SEQ ID NO:
- 14;
- administering the triple-fusion hESCs to the subject;
- and
- detecting fluorescence, bioluminescence, a nuclear signal from a PET reporter, or a combination thereof, emitted from the triple-fusion hESCs and any progeny of the triple-fusion hESCs.
8 more elements
A triple-fusion construct comprising:
- a firefly luciferase reporter gene (Fluc) having a nucleotide sequence selected from the group consisting of:
- SEQ ID NO:
- 1 and SEQ ID NO:3;
- a red fluorescent reporter gene (RFP) having a nucleotide sequence SEQ ID NO:
- 6;
- and a truncated Herpes Simplex Virus 1 thymidine kinase positron emission tomography (PET) reporter gene (HSV1-tTK) having a nucleotide sequence selected from the group consisting of:
- SEQ ID NO:
- 7 and SEQ ID NO:
- 14.
6 more elements
A method of monitoring stem cells comprising:
- providing stem cells;
- transducing the stem cells with a triple-fusion expression vector to produce triple-fusion stem cells, wherein the triple-fusion expression vector comprises a firefly luciferase reporter gene (Fluc) having a nucleotide sequence selected from the group consisting of:
- SEQ ID NO:
- 1 and SEQ ID NO:
- 3, a red fluorescent reporter gene (RFP) having a nucleotide sequence of SEQ ID NO:
- 6, and a truncated HSV1-tK positron emission tomography (PET) reporter gene (HSV1-tTK) having a nucleotide sequence selected from the group consisting of:
- SEQ ID NO:
- 7 and SEQ ID NO:
- 14;
- and
- detecting fluorescence, bioluminescence, a nuclear signal from a PET reporter, or a combination thereof, emitted from the triple-fusion stem cells and any progeny of the triple-fusion stem cells.
8 more elements
A method of monitoring human stem cells in a subject comprising:
- providing triple fusion human stem cells having a triple-fusion expression vector comprising a firefly luciferase reporter gene (Fluc) having a nucleotide sequence selected from the group consisting of:
- SEQ ID NO:
- 1 and SEQ ID NO:
- 3, a red fluorescent reporter gene (RFP) having a nucleotide sequence of SEQ ID NO:
- 6, and a truncated HSV1-tK positron emission tomography (PET) reporter gene (HSV1-tTK) having a nucleotide sequence selected from the group consisting of:
- SEQ ID NO:
- 7 and SEQ ID NO:
- 14;
- administering the triple-fusion human stem cells to the subject;
- and
- detecting fluorescence, bioluminescence, a nuclear signal from a PET reporter, or a combination thereof, emitted from the triple-fusion human stem cells and any progeny of the triple-fusion human stem cells.
8 more elements
Dependencies are inferred from the claim text.