Detection of RNA
OwnerWuPo Ma, Victor Lyamichev +6inventors
InventorsWuPo Ma, Victor Lyamichev, Michael Kaiser, Natalie E. Lyamichieva, Hatin Taysir Allawi, Andrew A. Lukowiak, James J. Schaefer, Andrew A. Lukowiak
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Abstract
The present invention provides novel cleavage agents and polymerases for the cleavage and modification of nucleic acid. The cleavage agents and polymerases find use, for example, for the detection and characterization of nucleic acid sequences and variations in nucleic acid sequences. In some embodiments, the 5′ nuclease activity of a variety of enzymes is used to cleave a target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.
Claims8
A composition comprising a nucleic acid sequence selected from the group consisting of 340, 345, 347, 350, 352, 358, 364, 366, 368, 373, 375, 379, 383, 387, 391, 395, 399, 401, 405, 407, 409, 411, 415, 417, 419, 423, 426, 428, 431, 435, 439, 443, 445, 447, 449, 452, 454, 455, 459, 463, 467, 471, 475, 481, 484, 495, 497, 499, 501, 505, 509, 513, 517, 521, 525, 529, 533, 537, 541, 543, 549, 552, 559, 563, 565, 567, 571, 573, 575, 577, 579, 581, 583, 585, 587, and 589 and the complements thereof.
A composition comprising a polypeptide encoded by a nucleic acid selected from the group consisting of SEQ ID NO:
- 340, 345, 347, 350, 352, 358, 364, 366, 368, 373, 375, 379, 383, 387, 391, 395, 399, 401, 405, 407, 409, 411, 415, 417, 419, 423, 426, 428, 431, 435, 439, 443, 445, 447, 449, 452, 454, 455, 459, 463, 467, 471, 475, 481, 484, 495, 497, 499, 501, 505, 509, 513, 517, 521, 525, 529, 533, 537, 541, 543, 549, 552, 559, 563, 565, 567, 571, 573, 575, 577, 579, 581, 583, 585, 587, and 589.
A method for producing an altered enzyme with improved functionality in a nucleic acid cleavage assay comprising:
- a) providing:
- i) a polypeptide comprising a sequence selected from the group consisting of SEQ ID NO:
- 340, 345, 347, 350, 352, 358, 364, 366, 368, 373, 375, 379, 383, 387, 391, 395, 399, 401, 405, 407, 409, 411, 415, 417, 419, 423, 426, 428, 431, 435, 439, 443, 445, 447, 449, 452, 454, 455, 459, 463, 467, 471, 475, 481, 484, 495, 497, 499, 501, 505, 509, 513, 517, 521, 525, 529, 533, 537, 541, 543, 549, 552, 559, 563, 565, 567, 571, 573, 575, 577, 579, 581, 583, 585, 587, and 589;
- ii) a nucleic acid encoding a polypeptide comprising a sequence selected from the group consisting of SEQ ID NO:
- 340, 345, 347, 350, 352, 358, 364, 366, 368, 373, 375, 379, 383, 387, 391, 395, 399, 401, 405, 407, 409, 411, 415, 417, 419, 423, 426, 428, 431, 435, 439, 443, 445, 447, 449, 452, 454, 455, 459, 463, 467, 471, 475, 481, 484, 495, 497, 499, 501, 505, 509, 513, 517, 521, 525, 529, 533, 537, 541, 543, 549, 552, 559, 563, 565, 567, 571, 573, 575, 577, 579, 581, 583, 585, 587, and 589;
- and
- iii) a nucleic acid test substrate;
- b) introducing one or more heterologous domains into said nucleic acid to produce an altered nucleic acid encoding an altered enzyme;
- c) contacting said altered enzyme and said polypeptide with said nucleic acid test substrate to produce cleavage products;
- and
- d) comparing cleavage products produced by said altered enzyme to cleavage products produced by said polypeptide.
A composition comprising a nucleic acid, said nucleic acid comprising sequence of SEQ ID NO:
- 543.
A composition comprising a polypeptide, said polypeptide comprising SEQ ID NO:
- 544.
Dependencies are inferred from the claim text.