CRISPR-assisted bioanalytical sensing of cytokines

Abstract

A vital component of innate and adaptive inflammatory host defenses is a soluble extracellular protein called cytokine, which regulates cell differentiation, growth, angiogenesis, and cell death, as well as development and repair processes that assist in re-establishing homeostasis. Cytokines, biomarkers for many diseases, are highly valuable across biology and clinical medicine. Their quantification can help with diagnosis and therapy by offering insights into pathological and physiological processes. Due to complicated cytokine networks and low concentration, cytokines are difficult to measure. Prominent conventional immunoassays require multiple steps for the loading of antibodies and samples, which makes detection laborious and time-consuming. Also, the sensitivity is low due to a lack of inherent signal amplification. The emergence of advanced multifarious technology like Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) may be able to meet the unmet demands of real-time, quick, and highly sensitive detection. CRISPR-Cas technology is a new approach to biosensing that is widely recognized for its exceptional precision. Recently, CRISPR-based diagnostic systems have undergone an advancement from genome editing to a clinically applicable diagnostic method for the rapid and highly sensitive identification of biomarkers, including DNAs/RNAs (circulating tumor DNAs or microRNAs), Ochratoxin A, proteins including calprotectin, and cytokines. Early diagnosis and prevention of chronic diseases can be greatly improved by the comprehensive characterization and timely analysis of cytokines using CRISPR-Cas technology. Since the finding of distinct endonuclease activity, this technology has significantly advanced the development of bioanalytical sensors. In particular, sophisticated disease diagnostic systems are developed using the multiplexed sensing capacities of CRISPR to address critical health problems. The following review explores the analytical tools and techniques that have been built on CRISPR-Cas technology for molecular detection, especially cytokines. It emphasizes the advantages, challenges, engineering strategies, and technological advancements of CRISPR-Cas systems in biosensing, intending to stimulate innovative concepts to tackle practical issues.