Screening artificial nucleic acids by particle display
AssigneeThe Regents of the University of California
InventorJohn Chaput
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Abstract
The invention provides xeno-nucleic acid particle display libraries, methods for identifying functional non-natural nucleic acid (XNA) aptamers using the particle display libraries, and compositions comprising XNA aptamers identified by screening candidate molecules using the xeno-nucleic acid particle display libraries.
Claims17
A method of producing a monoclonal xeno-nucleic acid aptamer particle (mXNAP) display library comprising:
- (a) providing a population of particles comprising a plurality of clonal template DNA molecules, each of which comprises a DNA coding region and a fixed sequence primer binding site;
- (b) ligating a self-priming stem-loop forming hairpin DNA molecule to the 3′ end of the template DNA molecules;
- (c) extending the 3′ end of the self-priming stem-loop in the presence of a polymerase capable of synthesizing an XNA from a DNA template and one or more XNA triphosphate molecules (xNTPs) to form a population of particles comprising clonal double stranded XNA-DNA hybrid molecules;
- (d) contacting the double stranded XNA-DNA display templates with a primer which anneals to the loop region of the stem-loop structure;
- and
- (e) extending the DNA primer using dNTPs and a DNA polymerase to displace the XNA portion of the XNA-DNA display templates to form a population of mXNAP display particles comprising a plurality of clonal nucleic acid molecules comprising a dsDNA region and a single-stranded XNA region.
A mXNAP display library comprising a population of display particles, wherein each display particle comprises a plurality of clonal nucleic acid molecules comprising a dsDNA region and a single-stranded XNA aptamer region.
A TNA aptamer, comprising a sequence selected from the group consisting of SEQ ID NO:1 to SEQ ID NO:3, a variant of SEQ ID NO:1 to SEQ ID NO:3, and a fragment comprising at least 20 nt of SEQ ID NO:1 to SEQ ID NO:3.
Dependencies are inferred from the claim text.