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US 8,298,764 B2Granted
Selection method for cell internalizing nucleic acids
AssigneeBoard of Regents, The University of Texas System
InventorsAndrew D. Ellington, Matthew Levy, Amy Yan, Ted Chitai Chu
Life of this patent
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Abstract
The present invention includes compositions and methods for contacting one or more cells with a random RNA-containing library, treating the contacted cells and with a denaturing agent or digestion with one or more nucleases, and extracting from the cells one or more internalized nucleic acids resistant to the nucleases or denaturants.
Claims22
A method for selecting RNA-containing molecules capable of being internalized, the method comprising:
- a) contacting a cell with a random library of RNA-containing molecules, wherein RNA-containing molecules capable of being internalized are internalized by the cell;
- b) exposing the contacted cells to one or more nucleases having RNase activity, wherein RNA-containing molecules that are not internalized by the cell are degraded by the nucleases;
- c) extracting total RNA from the contacted cells;
- and
- d) expanding the internalized RNA-containing molecules from the total RNA isolated from the cells,
- in selecting for RNA-containing molecules capable of being internalized.
3 more elements
A method for selecting stabilized RNA-containing molecules capable of being internalized, comprising:
- a) contacting one or more cells with a random nucleic acid library of RNA-containing molecules, the RNA-containing molecules comprising stabilized nucleic acids and containing constant sequence regions;
- b) washing the cells;
- c) exposing the cells to one or more nucleases having RNase activity;
- d) extracting total ribonucleic acids from the cells;
- e) expanding the RNA-containing molecules from the total ribonucleic acids;
- in selecting for RNA-containing molecules capable of being internalized;
- and
- f) repeating selection steps a)-e),
- wherein each subsequent round of selection comprises contacting the cells with the expanded nucleic acid pool iteratively for decreasing lengths of time.
6 more elements
Dependencies are inferred from the claim text.