Sequence specific degradation of single-stranded polynucleotides with CARD1 nuclease
AssigneeThe Rockefeller University
InventorsLuciano Marraffini, Jakob Rostol
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Abstract
Provided is an isolated or recombinantly expressed protein comprising the sequence of SEQ ID NO: 1 (CARD1), or an amino acid sequence that is at least 90% identical to the sequence of CARD1. Methods for producing and isolating the CARD1 protein are provided. Also provided are methods that include using CARD1, CaslO and RNA obtained from a biological sample, and a guide RNA targeted to an RNA polynucleotide that may be in the biological sample, and determining whether or not the CARD1 cleaves a reporter ssDNA or reporter ssRNA that is added to the sample, to determine the presence or absence of the RNA polynucleotide. Kits for use in the assay are also provided.
Claims2
A method for detecting a target RNA, wherein said method comprises (i) contacting a sample containing a target RNA with a protein that comprises SEQ ID NO:
- 1, a protein that comprises SEQ ID NO:
- 3, a guide RNA that is complementary to said target RNA and forms a complex with the protein that comprises SEQ ID NO:
- 3, and either a reporter single stranded DNA (ssDNA) or a reporter single stranded RNA (ssRNA), and (ii) detecting cleavage of the reporter ssDNA or the reporter RNA, wherein detection of said cleavage indicates the presence of the target RNA in the sample.
Dependencies are inferred from the claim text.