Methods for Identifying Fragile Histidine Triad (FHIT) Interaction and Uses Thereof
AssigneeThe Ohio State University Research Foundation
InventorsCarlo M. Croce, Francesco Trapasso
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Abstract
Provided herein are methods and compositions for the diagnosis, prognosis and treatment of a cancer associated disorder using the Fhit gene.
Claims21
An adenovirus being capable of isolating Fhit-His6, comprising an adenovirus carrying a FHIT-His6 cDNA.
A method of isolating exogenously over-expressed Fhit-His6, comprising using an adenovirus carrying a FHIT-His6 cDNA wherein the Fhit-His6 is isolated through the His tag.
A recombinant adenovirus carrying fragile histidine triad (Fhit) FHIT cDNA modified at its 3′ with a sequence encoding a histidine-six epitope tag (AdFHIT-His6).
A method for mediating an apoptotic process in at least one cell, comprising exposing the cell to a fragile histidine triad (Fhit) gene product in an amount sufficient to mediate the apoptotic process in the cell.
A method for inducing an apoptosis process in a cell, comprising exposing the cell to a fragile histidine triad (Fhit) gene product in an amount sufficient to cause generation of reactive oxygen species (ROS) in the cell.
A method for mediating an apoptotic process in at least one cell, comprising:
- exposing the cell to a sufficient amount of fragile histidine triad (Fhit) gene product to allow the Fhit to enter mitochondria in the cell and to interact with Fdxr protein in the cell and to cause an increase in Fdxr protein level that is associated with generation of ROS, and causing a change in the apoptotic process in the cell.
A method for preparing a wild-type FDXR (ferredoxin reductase) cDNA, comprising:
- PCR amplifying with primer sequences:
- 5′-CTgTTCCCAgCCATggCTTCgCgCTg-3′ (forward) [SEQ ID NO:
- 28], and 5′-TCAgTggCCCAggAggCgCAgCATC-3′ [SEQ ID NO:
- 29];
- and subcloning the amplification products pcDNA3.1/V5-HisTOPO TA vector.
A method for preparing a V5-tagged FDXR (ferredoxin reductase) cDNA, comprising:
- PCR amplifying with primer sequences:
- Forward:
- 5′-CTgTTCCCAgCCATggCTTCgCgCTg-3′ [SEQ ID NO:
- 30], reverse:
- 5′-gTggCCCAggAggCgCAgCATC-3′[SEQ ID NO:
- 31];
- and subcloning the amplification products pcDNA3.1/V5-HisTOPO TA vector.
A method for generating a recombinant adenovirus, comprising:
- preparing a recombinant adenovirus carrying the wild-type FHIT cDNA (AdFHIT);
- and generating a His-tagged FHIT cDNA using PCR with the following oligonucleotides:
- 5′-ACgTggATCCCTgTgAggACATgTCgTTCAgATTTggC-3′ (forward) [SEQ ID NO:
- 26], and 5′-TTgTggATCCTTATCAgTgATggTgATggTgATgCgATCCTCTCTgAAAgTAg ACCCgCAg-3′ [SEQ ID NO:
- 27].
-30. (canceled)
A method for identifying a correlation between Fhit expression and a disease or condition comprising identifying Fhit differentially expressed in a sample representative of the disease or condition compared to a normal sample.
-49. (canceled)
Dependencies are inferred from the claim text.