Enhanced guide nucleic acids and methods of use
AssigneeMammoth Biosciences, Inc.
InventorsWilliam Douglass WRIGHT, Benjamin Julius RAUCH, Aaron DELOUGHERY, James Paul BROUGHTON
Life of this patent
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Abstract
The present disclosure describes CRISPR/Cas proteins and guide RNA combinations that are capable of modifying nucleic acids at high temperatures. These compositions are especially useful for systems and methods of nucleic acid detection, including diagnostic devices.
Claims21
1-55. (canceled)
A composition comprising:
- a) a programmable nuclease or a nucleic acid encoding the programmable nuclease, wherein the programmable nuclease comprises an amino acid sequence that is at least 85% identical to SEQ ID NO:
- 1;
- and
- b) an engineered guide nucleic acid or DNA molecule encoding the engineered guide nucleic acid, wherein the engineered guide nucleic acid comprises a tracrRNA sequence that binds to the programmable nuclease and is at least 70% identical to any one of SEQ ID NO:
- 17-19, 41-42, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 88, 91, 94, 97, 100, 103, and 106, and wherein the tracrRNA sequence comprises a length of less than 140 linked nucleosides.
A composition comprising an engineered guide nucleic acid molecule configured to form a complex with a Cas protein, wherein said engineered guide nucleic acid molecule comprises:
- a spacer sequence exhibiting specific binding to a target gene;
- and
- a tracrRNA sequence for forming the complex with said Cas protein, wherein the tracrRNA sequence comprises at least 40 contiguous nucleobases of any one of SEQ ID NO:
- 17-19, 41-42, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 88, 91, 94, 97, 100, 103, and 106, and wherein the tracrRNA sequence comprises a length of less than 140 linked nucleosides.
A composition comprising an engineered guide nucleic acid molecule configured to form a complex with a Cas protein, wherein said engineered guide nucleic acid molecule comprises:
- a spacer sequence exhibiting specific binding to a target gene;
- and
- a tracrRNA sequence for forming the complex with said Cas protein, wherein the tracrRNA sequence is at least 70% identical of any one of SEQ ID NO:
- 17-19, 41-42, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 88, 91, 94, 97, 100, 103, or 106;
- wherein the length of the tracrRNA sequence is less than 140 linked nucleosides.
Dependencies are inferred from the claim text.