Programmable nucleic acid detection with CRISPR-Cas12a and CRISPR-Cas13a/d

AbstractThe CRISPR (Clustered Interspaced Short Palindromic Repeats) system has an impact on the immune defense of prokaryotes, like bacteria and archaea. It uses RNA-guided nucleases to target and break down foreign genomes through already acquired sequence memory from the same invading genome. This process has three main steps: spacer acquisition, expression, and interference. Scientists group CRISPR into classes, Cas9, Cas12, and Cas13 proteins playing crucial roles in different CRISPR types. Cas12a and Cas13a/d have become essential tools to detect nucleic acids allowing for very accurate and specific diagnoses. New developments have combined CRISPR-based diagnostics with microfluidic systems, ways to boost signals, and platforms that can detect multiple things at once such as SHERLOCK can act through Cas13 recognition of target nucleic acid species with high sensitivity and specificity and DETECTR which relies on the collateral activity of Cas12a protein RNA. These systems offer quick, cheap, and portable ways to spot viral and bacterial pathogens. Yet, problems like unintended effects and technical hurdles in clinical use still exist. Ongoing improvements in CRISPR technology are likely to make it more precise and useful in various fields, including molecular diagnostics and pathogen detection. Integrating CRISPR with biosensors and artificial intelligence based detection platforms has attracted much attention as highly specific and sensitive sensors with easily programmable and device- independent capability in nature. It will make CRISPR available for many upcoming research works.