A sequence-specific plasmonic loop-mediated isothermal amplification assay with orthogonal color readouts enabled by CRISPR Cas12a

Abstract

CRISPR Cas12a enables a sequence-specific plasmonic LAMP assay with dual complementary color readouts.

Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities

“…However, instead of only a red to blue color shift as a read-out, two reaction tubes where used for orthogonal color readout. This method allows quantification in a larger concentration range than usual ( Zhou et al, 2020 ).…”

CRISPR‐Cas12a System for Biosensing and Gene Regulation

“…The ssDNA substrates of collateral cleavage can also be stabilizers preventing salt‐induced aggregation of bare AuNPs. Thus, a sequence‐specific plasmonic sensing platform exhibits dual‐color readouts, which can minimize the influence of environmental factors causing false positives or negatives, due to orthogonality between the dual‐color channels [33b] …”

Amplified detection of nucleic acids and proteins using an isothermal proximity CRISPR Cas12a assay

“…[1][2][3] Recent advances in microbial clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (CRISPR-Cas) enzymes offer exciting opportunities for developing novel isothermal nucleic acid amplication assays that are both sensitive and specic. [4][5][6][7][8][9][10][11][12][13][14][15][16][17][18] In particular, the combination of recombinase polymerase amplication (RPA) with the unique target-binding induced indiscriminative singlestranded DNase (ssDNase) activity of Cas12 and Cas13 has led to the development of a series of isothermal assays, such as specic high-sensitivity enzymatic reporter unlocking (SHERLOCK) [4][5][6] and DNA endonuclease-targeted CRISPR trans reporter (DETECTR). 7 We have also introduced an isothermal plasmonic CRISPR assay by integrating Cas12a with loopmediated isothermal amplication (LAMP) and plasmonic nanoparticles.…”

“…7 We have also introduced an isothermal plasmonic CRISPR assay by integrating Cas12a with loopmediated isothermal amplication (LAMP) and plasmonic nanoparticles. 8 So far, the detection of each specic genetic marker requires the existence of a protospacer-adjacent motif (PAM) domain and the design of a specic CRISPR RNA (crRNA). Herein, we introduce a new isothermal amplication assay principle, where target recognition is achieved through proximity hybridization [17][18][19][20][21] rather than crRNA binding.…”