CRISPR-Mediated Cleavage of Enzyme-Oligonucleotide Conjugates for Detection of Target Analytes

NU 2021-068

INVENTORS

• Chad Mirkin*
• Devleena Samanta
• Sasha Ebrahimi

SHORT DESCRIPTION

This technology leverages CRISPR-mediated cleavage of enzyme-oligonucleotide conjugates to enable sensitive, colorimetric detection of target analytes without the need for specialized equipment or stringent temperature conditions.

BACKGROUND

Current diagnostic platforms, such as PCR and ELISA, often require complex, multi-step processes, specific temperature conditions, and advanced instrumentation, limiting their use to centralized facilities. This invention addresses these limitations by providing a simplified, point-of-care diagnostic solution that can detect nucleic acid and non-nucleic acid analytes efficiently and effectively.

ABSTRACT

Northwestern researchers have developed a novel diagnostic strategy utilizing CRISPR-mediated cleavage of surface-bound enzyme-oligonucleotide conjugates. Upon the presence of a target analyte, a nucleic acid sequence binds to a CRISPR RNAse or DNAse, such as Cas13 or Cas12, pre-complexed with a guide RNA. This interaction activates the CRISPR enzyme, leading to the cleavage of the conjugates and the release of enzymes into solution. The subsequent treatment with an enzyme substrate produces a chromogenic, fluorogenic, or luminescent signal, which is directly proportional to the target concentration. This approach facilitates rapid and routine detection of viral and bacterial infections, disease biomarkers, and monitoring of disease progression or therapeutic response.

APPLICATIONS

• Detecting nucleic-acid and non-nucleic acids in solution and clinical samples
• Quantifying levels of analytes in serum

ADVANTAGES

• Specialized equipment not necessary
• No specific temperature requirement
• Signal amplification without target amplification
• Detectable by the naked eye in some cases
• Simple workflow for at-home or point-of-care use

IP STATUS
Issued US Patent US20240150817A1

PUBLICATION

Samanta, D., Ebrahimi, S.B., Ramani, N., Mirkin, C.A. Enhancing CRISPR-CAS-Mediated Detection of Nucleic Acids and Non-nucleic Acid Targets Using Enzyme-Labeled Reporters. J. Am. Chem. Soc. 2022, 144, 35, 16310-16315.