CRISPR-Cas Endonuclease Chain Reaction: a revolution in point-of-care genetic testing

CRISPR-Cas systems act as molecular detectives that get activated upon encountering genetic material from pathogens or specific DNA mutations. While holding great promise for genetic testing, they face a key limitation, as they require larger amounts of genetic material than typically found in test samples, such as nasal or oral swabs. Current solutions require complex pre-amplification steps, unsuitable for point-of-care or self-diagnosis settings. Our groundbreaking innovation, CRISPR-Cas Endonuclease Chain Reaction (ECR), or "Easier," provides an original solution by not amplifying genetic material, but instead, triggering a chain reaction of CRISPR-Cas activity. Further improvements to ECR allow to test multiple diseases and/or in multiple patients simultaneously. ECR aims to democratize and decentralize genetic testing, empowering individuals with accurate genetic information, free from traditional laboratory constraints. This is the transformative future we aspire to deliver.

ECR is a patented nucleic-acid detection method based on CRISPR-Cas technology that allows fully customizable, precise and sensitive detection of virtually any nucleic acid sequence of interest at point of-care. By adapting it to a portable sequencing readout, ECR will further acquire ultrasensitive and multiplexing capabilities, allowing bedside analysis of pathogen or genetic-marker panels, while also being competitive for massive genetic screening in centralized facilities.

CRISPR-Cas nucleic-acid detection methods rely on Cas12 and Cas13 ribonucleoparticles (RNPs) that, upon recognition and cleavage of a specific target sequence (DNA or RNA, respectively), acquire a collateral endonuclease activity that degrades detectable reporter molecules. However, this activity is insufficient for direct diagnosis and currently requires pre-amplification of the target sequence. In ECR, amplifier molecules designed to release targets for a second CasRNP upon collateral activity, generate a chain reaction of Cas activation that exponentially amplifies the signal, ultimately achieving full degradation of reporter substrates. This process results in a 1000x increase in sensitivity, only limited by the eventual appearance of background signal. Coupled to additional improvements, such as non targeting decoys and barcode-ligation sequencing, ECR acquires single-nucleotide precision, single-molecule counting sensitivity and high multiplexing capacity.