Target-specific compomers and methods of use
View Patent ↗Provided herein are libraries of nucleic acid species each comprising a transcription unit having a promoter region operatively linked to a coding sequence. The coding sequence of each nucleic acid species encodes a RNA cleavage substrate comprising a unique compomer species and a cleavage site. Each compomer species has a molecular mass distinguishable from the molecular mass of other compomer species in the library, and cleavage at a cleavage site releases a polynucleotide comprising the compomer species from the RNA cleavage substrate.
1. A method for identifying the presence or absence of a target nucleic acid in a sample, which comprises:
(a) contacting a sample with a target detection reagent that forms a reagent:target complex when target is present in the sample, wherein:
(i) the target detection reagent comprises a transcription unit comprising a promoter region operatively linked to a coding sequence;
(ii) the coding sequence encodes a RNA transcription product;
(b) transcribing a RNA transcription product from the coding sequence in a complex present after (a), wherein the RNA transcription product comprises a compomer consisting of one, two or three nucleotide types, and a terminal nucleotide sequence consisting of one or more nucleotide types not included in the compomer;
(c) cleaving the RNA transcription product thereby generating a cleavage product, wherein the 3′ terminus of the compomer is at the 3′ terminus of the cleavage product; and
(d) detecting the presence or absence of the cleavage product that includes the compomer, whereby the presence or absence of the target nucleic acid is identified based on the presence or absence of the cleavage product.
2. The method of claim 1 , wherein the cleavage product consists of the compomer.
3. The method of claim 1 , wherein the RNA transcription product consists of the compomer and a terminal nucleotide sequence common to one or more other RNA transcription products.
4. The method of claim 3 , wherein the nucleotides in the compomer and terminal nucleotide sequence are selected from the group consisting of adenine (A), thymine (T), guanine (G), and cytosine (C).
5. The method of claim 1 , wherein the RNA transcription product consists of the compomer and the presence or absence of the compomer is identified without cleaving the RNA transcription product.
6. The method of claim 1 , wherein the presence or absence of the cleavage product is identified by mass spectrometry.
7. The method of claim 5 , wherein the presence or absence of the cleavage product is identified in a mass range of about 2500 Daltons and about 10000 Daltons.
8. The method of claim 1 , wherein the RNA transcription product is cleaved by a process selected from the group consisting of a chemical, a physical, and an enzymatic process.
9. The method of claim 1 , wherein the promoter region is selected from the group consisting of a bacterial, a bacteriophage, a consensus, a viral and a eukaryotic promoter region.
10. The method of claim 1 , wherein the promoter region is a bacteriophage promoter region selected from the group consisting of a T7, an SP6, and a T3 promoter region.
11. The method of claim 1 , wherein the target nucleic acid is selected from the group consisting of a source of genetic variation among members of the same species and a chemically modified nucleic acid molecule.
12. The method of claim 1 , wherein the target detection reagent is one of a plurality of target detection reagents, and wherein RNA transcription product comprises a unique compomer species.