Universal paper-based genome detection using a universal rna toehold switch and crispr
AssigneeThe Research Foundation for The State University of New York
InventorsMehmet Yigit, Mahera Kachwala
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Abstract
Swift and accurate on-site identification of genomes of pathogens is crucial to curb its spread and contamination. In embodiments disclosed herein, a synthetic biology cascade reaction is presented on a paper substrate using CRISPR-Cas12a and recombinase polymerase amplification (RPA), enabling the programming of a standard toehold RNA switch for a genome of choice through insertion of meticulously designed an oligonucleotide (probe AB).
Claims20
A method for visually detecting a Salmonella genome on a paper substrate, said method comprising:
- a) applying a sample that might contain the genome to the paper substrate;
- b) performing a cascade reaction including a CRISPR-Cas12a reaction utilizing a CRISPR-Cas12a-crRNA complex on the paper substrate;
- c) conducting a toehold RNA cascade reaction on the paper substrate;
- and d) visually detecting the presence of Salmonella genome based on the results of steps a), b) and c).
A paper-based diagnostic device for visually detecting Salmonella genome, comprising:
- a) a paper substrate containing freeze-dried components for a cascade reaction including CRISPR-Cas12a reaction and toehold RNA cascade reaction;
- b) a Cas12a-crRNA complex wherein the crRNA is designed complementary to bind Salmonella genome amplicons;
- c) a toehold switch linked to a LacZ gene;
- and d) a chromogenic substrate for β-galactosidase.
A method for visually detecting a genome comprising:
- a) applying a sample that might contain the genome to a paper substrate;
- b) performing a cascade reaction including a CRISPR-Cas12a reaction on the paper substrate;
- c) conducting a toehold RNA cascade reaction on the paper substrate;
- and d) visually detecting the presence of a genome based on the results of steps a), b), and c).
Dependencies are inferred from the claim text.