Crispr effector system based diagnostics for hemorrhagic fever detection
AssigneesThe Broad Institute, Inc., President and Fellows of Harvard College
InventorsPardis Sabeti, Cameron Myhrvold, Catherine Amanda Freije, Anna Elizabeth Lachenauer, Kayla Grace Barnes
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Abstract
The embodiments disclosed herein utilize RNA targeting effectors to provide a robust CRISPR-based diagnostic for hemorrhagic fever virus applications. Embodiments disclosed herein can differentiate between hemorrhagic fever viruses that present with similar symptoms, as well as between strains of a hemorrhagic fever virus.
Claims68
A nucleic acid detection system for detecting the presence of hemorrhagic fever viruses in a sample comprising:
- a CRISPR system comprising an effector protein and one or more guide molecules designed to bind to one or more corresponding target molecules of one or more hemorrhagic fever viruses;
- and
- an RNA-based masking construct.
A method for detecting viral nucleic acid in a sample comprising:
- amplifying the sample nucleic acid;
- combining the sample with an RNA effector protein, one or more guide molecules according to SEQ ID NOs:
- 80, 87-92, 109-126, 139-156, 159-172, 207-228, 249-281, 329-366, and 393-416, and an RNA-based masking construct, wherein the one or more guide molecules are designed to bind to corresponding virus specific target molecules;
- activating the RNA effector protein via binding of the one or more guide molecules to the one or more virus-specific target molecules, wherein activating the RNA effector protein results in modification of the RNA-based masking construct such that a detectable positive signal is produced;
- and
- detecting the signal, wherein detection of the signal indicates the presence of a hemorrhagic fever virus;
- and
- wherein the method does not include the step of extracting nucleic acid from the sample.
A kit for detecting viral nucleic acids in a sample, comprising
- nucleic acid amplification reagents;
- a CRISPR system comprising an effector protein and one or more of the guide RNAs according to SEQ ID NO:
- 80, 87-92, 109-126, 139-156, 159-172, 207-228, 249-281, 329-366, and 393-416, wherein the guide RNAs are designed to bind to corresponding target molecules;
- an RNA-based masking construct;
- and
- one or more lateral flow strips.
Dependencies are inferred from the claim text.