Methods for improving and managing nucleoside reverse-transcriptase inhibition based treatment
AssigneeAmikana, Biologics
InventorsPablo Gluschankof, Christele Perrin-East
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Abstract
A transformed yeast cell includes: a nucleic acid sequence coding for a reverse transcriptase; a reverse transcription indicator; a nucleic acid sequence coding for deoxycytidine kinase (dCK); and at least one nucleic acid sequence coding for nucleoside transporter. Methods for screening a compound with the ability to inhibit reverse transcription and for predicting the sensitivity of a reverse transcriptase to a NRTI compound, particularly a reverse transcriptase derived from a virus infecting a subject using the transformed Yeast cell are also described.
Claims6
A method for determining the sensitivity of a reverse transcriptase to a nucleoside reverse transcriptase inhibitor (NRTI) compound selected from the group consisting of Zidovudine, Didanosine, Stavudine, Tenofovir, Zalcitabine and combinations thereof, said method comprising the steps of:
- i. providing recombinant yeast cells comprising:
- a nucleic acid sequence coding for a reverse transcriptase,
- a reverse transcription indicator enabling selection of transformed yeast cells in which reverse transcription occurs,
- a nucleic acid sequence coding for deoxycytidine kinase (dCK), and
- a nucleic acid sequence coding for the nucleoside transporter OAT1,
- ii. contacting said yeast cells separately with and without said NRTI compound,
- iii. culturing the yeast cells in a selective medium,
- iv. determining the growth of the yeast cells contacted with or without said NRTI compound, and
- v. determining therefrom if the reverse transcriptase to be studied is sensitive or resistant to said NRTI compound, wherein a decrease in the growth of the yeast cells contacted with said NRTI compound of at least 30% relative to the growth of the yeast cells that are not contacted with said NRTI compound is indicative of reverse transcriptase sensitivity to said NRTI compound.
Dependencies are inferred from the claim text.