Back to results
US 2014/0112871 A1Published application
Nucleic acid probes and methods of using the same
AssigneeBioVentures, LLC
InventorsAlexei G. Basnakian, Yevgeniy Apostolov
Life of this patent
Estimating the term…
Abstract
The present invention provides a composition comprising a fluorescent nucleic acid probe that can be cleaved by enzymatic and non-enzymatic means, and methods of using the same. Advantageously, the nucleic acid probe may be used to detect and quantify nucleic acid cleavage in vitro, in situ, ex vivo, and in vivo.
Claims20
A method for detecting nucleic acid cleavage in a sample, the method comprising:
- (i) providing a nucleic acid probe comprising two components, A1-A2-A3 and A5-A6-A7, and optionally a third component, A4;
- wherein
- (a) A1 and A7 are fluorophores,
- (b) A2 and A6 are each a single-stranded nucleic acid about 1 to about 50 nucleotides in length that are not complementary to the other nor to A3, A4, or A5, and at least one of A2 or A6 comprises a nuclease recognition sequence,
- (c) A3 and A5 are each a single-stranded nucleic acid about 2 to about 80 nucleotides in length and comprise a region complementary to the other, such that A3 and A5 associate when A1-A2-A3 and A5-A6-A7 are not cleaved and disassociate when at least one of A1-A2-A3 or A5-A6-A7 is cleaved,
- (d) A4 is a single-stranded nucleic acid comprising about 4 nucleotides to about 50 nucleotides in length that, when present, connects and forms a loop structure between A3 and A5,
- whereby the fluorescence of A1 and A7 is quenched when A3 and A5 associate and is detectable when A3 and A5 disassociate;
- (ii) contacting the sample with the nucleic acid probe and detecting fluorescence in the sample, whereby fluorescence in a sample indicates nucleic acid cleavage of at least one component of the nucleic acid probe and dissociation of A3 and A5;
- and
- (iii) optionally, quantifying the fluorescence in the sample.
7 more elements
A method for detecting nucleic acid cleavage in a subject, the method comprising:
- (i) administering to a subject a nucleic acid probe comprising two components, A1-A2-A3 and A5-A6-A7, and optionally a third component, A4;
- wherein
- (a) A1 and A7 are near-infrared fluorophores,
- (b) A2 and A6 are each a single-stranded nucleic acid about 1 to about 50 nucleotides in length that are not complementary to the other nor to A3, A4, or A5, and at least one of A2 or A6 comprises a nuclease recognition sequence,
- (c) A3 and A5 are each a single-stranded nucleic acid about 2 to about 80 nucleotides in length and comprise a region complementary to the other, such that A3 and A5 associate when A1-A2-A3 and A5-A6-A7 are not cleaved and disassociate when at least one of A1-A2-A3 or A5-A6-A7 is cleaved,
- (d) A4 is a single-stranded nucleic acid comprising about 4 nucleotides to about 50 nucleotides in length that, when present, connects and forms a loop structure between A3 and A5,
- whereby the fluorescence of A1 and A7 is detectable when A3 and A5 disassociate and is quenched when A3 and A5 associate;
- and
- (ii) performing a signal acquisition scan on the subject to detect the fluorescence signal, whereby fluorescence indicates nucleic acid cleavage of at least one component of the nucleic acid probe and dissociation of A3 and A5.
6 more elements
A composition comprising a nucleic acid probe, the nucleic acid probe comprising two components:
- A1-A2-A3 and A5-A6-A7, wherein:
- (a) A1 and A7 are fluorophores,
- (b) A2 and A6 are each a single-stranded nucleic acid about 1 to about 50 nucleotides in length that are not complementary to the other nor to A3 or A5, and at least one of A2 or A6 comprises a nuclease recognition sequence,
- (c) A3 and A5 are each a single-stranded nucleic acid about 2 to about 80 nucleotides in length and comprise a region complementary to the other, such that A3 and A5 associate when A1-A2-A3 and A5-A6-A7 are not cleaved and disassociate when at least one of A1-A2-A3 or A5-A6-A7 is cleaved,
- whereby the fluorescence of A1 and A7 is quenched when A3 and A5 associate and is detectable when A3 and A5 disassociate.
2 more elements
Dependencies are inferred from the claim text.