CRISPR/Cas9‐Assisted Microrobots for Fast and Ultrasensitive “On‐The‐Fly” Next‐Generation DNA Detection

Abstract Fast, sensitive, and selective nucleic acids detection is essential for applications in clinical diagnostics, food safety, and environmental monitoring. Hence, in this study, we introduce a novel motion‐assisted CRISPR/Cas9 microrobots‐based biosensor, consisting of CRISPR/Cas9 functionalized on gold microrobots (Au–MRs) forming CRISPR/Cas9@Au–MRs, which exhibit self‐propulsion leading to rapid and ultrasensitive detection of target DNA. The proposed system achieved the limit of detection in low fM DNA concentration, enabling detection across a wide dynamic range within only 5 min, which is significantly faster than any previously reported systems. Further, this self‐propelled system removes the need for complicated sample preparation, enabling real‐time detection by using a minimal amount of sample. Unlike conventional CRISPR biosensors, this platform utilizes “on‐the‐fly” motion‐enhanced signal recovery, ensuring superior specificity and sensitivity in diverse biological environments. Importantly, the presented system shows exceptional analytical performance in biological samples, highlighting its potential for real‐world sensing applications. Overall, the motion‐assisted CRISPR/Cas9@Au–MRs biosensor represents evolutionary progress in nucleic acids detection, providing a fast, reliable platform that can be easily scalable for applications in disease detection, biosafety and environmental detection.