Advancements in CRISPR-Based Biosensing for Next-Gen Point of Care Diagnostic Application

Abstract

With the move of molecular tests from diagnostic labs to on-site testing becoming more common, there is a sudden rise in demand for nucleic acid-based diagnostic tools that are selective, sensitive, flexible to terrain changes, and cost-effective to assist in point-of-care systems for large-scale screening and to be used in remote locations in cases of outbreaks and pandemics. CRISPR-based biosensors comprise a promising new approach to nucleic acid detection, which uses Cas effector proteins (Cas9, Cas12, and Cas13) as extremely specialized identification components that may be used in conjunction with a variety of readout approaches (such as fluorescence, colorimetry, potentiometry, lateral flow assay, etc.) for onsite analysis. In this review, we cover some technical aspects of integrating the CRISPR Cas system with traditional biosensing readout methods and amplification technologies such as polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), and recombinase polymerase amplification (RPA) and continue to elaborate on the prospects of the developed biosensor in the detection of some major viral and bacterial diseases. Within the scope of this article, we also discuss the recent COVID pandemic and the numerous CRISPR biosensors that have undergone development since its advent. Finally, we discuss some challenges and future prospects of CRISPR Cas systems in point-of-care testing.

RNA-Based Sensor Systems for Affordable Diagnostics in the Age of Pandemics

“…However, antibody tests are often cheaper, quicker, and less sophisticated. They are essential in determining immunological state and prior infection, something that RNA-based sensors are not able to do. , …”

“…Hand-held cell-free systems can come in a wide range of different designs, ranging from microfluidic to single reaction chamber ones. They are generally easier to develop compared to paper-based because they have fewer components than paper-based systems and can thus be easily designed to be compatible with a wide range of reactions and analyses, hand-held cell-free systems are more versatile when it comes to adapting to small and portable dimensions for diagnosis and analysis . Example microfluidic-based systems range from simple ones, such as lateral flow assays (i.e., antibody tests), to more complex designs that work with smartphones to process the data they generate.…”

“…They are essential in determining immunological state and prior infection, something that RNA-based sensors are not able to do. 110,111 Cell-free systems have the advantage of having a simple nature. 93 They can easily be stored by freeze-drying and, when needed, could be activated with rehydration.…”

Recent Progress in Prompt Molecular Detection of Exosomes Using CRISPR/Cas and Microfluidic‐Assisted Approaches Toward Smart Cancer Diagnosis and Analysis

“…In contrast, liquid biopsy is limited to detecting genetic material, offering fewer options for direct disease management. [148][149][150][151] Cost-effectiveness is another area where CRISPR technology shines. Liquid biopsy, due to the need for specialized equipment and extensive sequencing, can be relatively expensive.…”

“…It can edit genes, correct mutations, and even activate or suppress specific genetic functions, making it a versatile tool for both diagnosis and potential therapeutic interventions. In contrast, liquid biopsy is limited to detecting genetic material, offering fewer options for direct disease management [148–151] . Cost‐effectiveness is another area where CRISPR technology shines.…”