COMPOSITION AND METHODS FOR DETECTING ADENOSINE MODIFICATIONS
First Claim
1. A method for detecting modified adenosine in a target ribonucleic acid (RNA) comprising contacting the target RNA with an adenosine deaminase enzyme to generate a target RNA with deaminated adenosines and sequencing the target RNA with deaminated adenosines;
- wherein any modified adenosine in the target RNA is read as an adenosine in the sequencing of the target RNA with deaminated adenosines.
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Abstract
The current disclosure relates to methods, compositions and kits for detecting modified adenosine in a target RNA molecule. Aspects relate to a method for detecting modified adenosine in a target ribonucleic acid (RNA) comprising contacting the target RNA with an adenosine deaminase enzyme (adenosine deaminase, RNA-specific) to generate a target RNA with deaminated adenosines and sequencing the target RNA with deaminated adenosines; wherein the modified adenosine is detected when the nucleotide sequence is adenosine.
19 Citations
76 Claims
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1. A method for detecting modified adenosine in a target ribonucleic acid (RNA) comprising contacting the target RNA with an adenosine deaminase enzyme to generate a target RNA with deaminated adenosines and sequencing the target RNA with deaminated adenosines;
- wherein any modified adenosine in the target RNA is read as an adenosine in the sequencing of the target RNA with deaminated adenosines.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57)
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58. A kit comprising:
- an adenosine deaminase enzyme and instructions for detecting modified adenosines in target RNA.
- View Dependent Claims (59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72)
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73. A method for detecting modified adenosine in a target ribonucleic acid (RNA) comprising contacting a double-stranded nucleic acid molecule comprising the target RNA with ADAR to generate a target RNA with deaminated adenosines and sequencing the target RNA with deaminated adenosines;
- wherein the modified adenosine is detected when the sequence of the target RNA with deaminated adenosines is adenosine.
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74. A method for detecting modified adenosine in a target ribonucleic acid (RNA) comprising:
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a) providing a target RNA; b) generating a DNA strand that is complementary with the target RNA and hybridizing the complementary DNA strand with the target RNA to generate a RNA-DNA duplex comprising the target RNA; c) contacting the RNA-DNA duplex with ADAR to generate target RNA with deaminated adenosines; and d) contacting the RNA-DNA duplex with a DNA digesting agent; and e) sequencing the target RNA with the deaminated adenosines.
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75. A method for detecting modified adenosine in a target ribonucleic acid (RNA) comprising:
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a) providing a target RNA; b) generating a DNA strand that is complementary with the target RNA and hybridizing the complementary DNA strand with the target RNA to generate a RNA-DNA duplex comprising the target RNA; c) contacting the RNA-DNA duplex with ADAR to generate target RNA with deaminated adenosines; and d) contacting the RNA-DNA duplex with a DNA digesting agent; e) repeating steps b, c, and d one or more times; and f) sequencing the target RNA with the deaminated adenosines.
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76. A method for detecting modified adenosine in a target ribonucleic acid (RNA) comprising:
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a) providing a target RNA; b) generating an RNA strand with modified adenosine that is complementary with the target RNA to generate an RNA-RNA duplex comprising the target RNA; c) contacting the RNA-RNA duplex with ADAR to generate target RNA with deaminated adenosines; and d) sequencing the target RNA with the deaminated adenosines.
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Specification