Volume 5 Number 1 2026
CRISPR-Cas13 Paper Biosensors for Point-of-Care Genotyping of Thalassemia Mutations: Mechanisms, Isothermal Coupling, and Translational Readiness
Mohammad Lutfor Rahman 1*, Md Shamsuddin Sultan Khan 2*
Biosensors and Nanotheranostics 5 (1) 1-8 https://doi.org/10.25163/biosensors.5110998
Submitted: 03 October 2026 Revised: 20 November 2026 Accepted: 29 November 2026 Published: 01 December 2026
Thalassemia is among the most common inherited monogenic disorders worldwide, and its heaviest burden falls on regions where centralized molecular testing is hardest to sustain. Accurate carrier screening and allelic discrimination remain the backbone of prevention, yet the reference methods that deliver them, polymerase chain reaction, reverse dot-blot, multiplex ligation-dependent probe amplification, and sequencing, depend on instruments, cold chains, and trained staff that many endemic settings cannot guarantee. This review examines how CRISPR-Cas13 paper biosensors might close that gap. We trace the enzymology of the Type VI effector, whose crRNA-guided recognition of single-stranded RNA unleashes a high-turnover collateral trans-cleavage that converts one binding event into thousands of cleaved reporters, and we show how a single engineered mismatch in the guide can sharpen that activity into single-nucleotide allelic resolution for mutations such as CD41/42, IVS-1-110, and codon 39. We then follow the full assay cascade, from rapid whole-blood lysis through isothermal recombinase polymerase amplification and T7 in vitro transcription to a lateral-flow dipstick on which a severed reporter is read as a visible band by the naked eye. Reagent lyophilization with disaccharide lyoprotectants is considered as the step that frees the platform from refrigeration. Across hematologic, oncologic, and infectious-disease applications, reported concordance with laboratory standards is high, though most thalassemia work remains preclinical. We argue that simplified extraction, one-pot contamination control, and robust multiplexing are the decisive hurdles between a promising chemistry and a deployable, REASSURED-aligned diagnostic. On current evidence the chemistry is mature, whereas thalassemia-specific clinical validation across endemic populations remains the critical unfinished step.
Keywords: CRISPR-Cas13; SHERLOCK; thalassemia; point-of-care testing; lateral-flow assay; recombinase polymerase amplification; single-nucleotide discrimination; collateral cleavage
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