Split CRISPR/Cas systems: Pioneering solutions for molecular diagnostics challenges

Split CRISPR/Cas systems may achieve significant improvements in sensitivity for femtomolar-level biomarkers.

• Split-activation strategies enhance analytical performance without the need for pre-amplification.
• Two key approaches include split activator-mediated Cas systems for sensitive RNA detection and split crRNA architectures for analyzing multiple nucleic acids.
• These innovations maintain superior single-base discrimination while improving detection capabilities.
• Applications include clinical diagnostics and on-site monitoring, though challenges such as background noise and cost remain.
• Future developments may focus on customized Cas variants and integrated platforms for improved diagnostics.

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