Composition and Methods of a Nuclease Chain Reaction for Nucleic Acid Detection

Tech ID: 31940 / UC Case 2020-120-0

Patent Status

Patent Pending

Brief Description

This invention leverages the nuclease activity of CRISPR proteins for the direct, sensitive detection of specific nucleic acid sequences. This all-in-one detection modality includes an internal Nuclease Chain Reaction (NCR), which possesses an amplifying, feed-forward loop to generate an exponential signal upon detection of a target nucleic acid.

Cas13 or Cas12 enzymes can be programmed with a guide RNA that recognizes a desired target sequence, activating a non-specific RNase or DNase activity. This can be used to release a detectable label. On its own, this approach is inherently limited in sensitivity and current methods require an amplification of genetic material before CRISPR-base detection. 

Suggested uses

All nucleic acid diagnostics and detection, both DNA and RNA.

Advantages

Current methods of programming Cas13 or Cas12 enzymes to release a detectable label require an amplification of genetic material before CRISPR-base detection and are inherently limited in sensitivity. In these existing scenarios, a guide RNA recognizes a desired target sequence, activating a non-specific RNase or DNase activity.

In contrast, this invention includes an all-in-one detection modality with an internal Nuclease Chain Reaction (NCR) and an amplifying, feed-forward loop. The technology generates an exponential signal upon detection of a target nucleic acid.

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Inventors

  • Savage, David Frank

Other Information

Keywords

nucleic acid, nuclease chain reaction, NCR, Cas13, Cas12

Categorized As