CRISPR biosensing with lateral flow assays for point of care Diagnostics: Overcoming commercial development challenges

The combination of lateral flow assays (LFAs) with CRISPR biosensing technologies offers a new frontier in point-of-care (POC) diagnostics. LFAs are widely used for their simplicity, rapid results, and ease of use, enabling detection of various targets including proteins and nucleic acids. However, traditional LFAs often struggle with sensitivity, especially in detecting viral nucleic acids like SARS-CoV-2. CRISPR biosensing systems, particularly those using Cas12 and Cas13 effectors, enhance LFA performance by providing highly sensitive and specific detection of nucleic acids. These effectors, upon recognizing a target sequence, trigger collateral cleavage activity, amplifying the detection signal. This approach has been used in diagnostic tools such as DETECTR (Cas12a) and SHERLOCK (Cas13a), which have shown PCR-level sensitivity and specificity for detecting viral RNA, including SARS-CoV-2, with faster and more cost-effective results. By integrating CRISPR technologies into LFAs, these hybrid systems are poised to offer powerful solutions for sensitive, low-cost, and user-friendly diagnostics, especially in resource-limited settings. This review summarizes the recent advancements in CRISPR-based LFAs, exploring their mechanisms, advantages, and limitations. In addition, this review also outlines the commercial development challenges and discussed the future prospects. • CRISPR based LFAs offers a new frontier in POC diagnostics. • CRISPR based LFAs represents a significant commercial opportunity. • CRISPR based LFAs is able to realize multiplex detection without preamplification.