Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities

van Dongen, Jeanne E.; Berendsen, Johanna T.W.; Steenbergen, Renske D.M.; Wolthuis, Rob M.F.; Eijkel, Jan C.T.; Segerink, Loes I.

With the trend of moving genomic test from (medical) laboratories to on-site testing, there is a need for nucleic acid based diagnostic tools combining the sensitivity, specificity and flexibility of established genomic diagnostics with the ease, cost effectiveness and speed of isothermal amplification methods. A promising new genomic sensing method is Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated nuclease (Cas)-based sensing. In this method Cas effector proteins are used as highly specific sequence recognition elements that can be combined with many different read-out methods for on-site point-of-care testing. This review covers the technical aspects of integrating CRISPR/Cas technology in miniaturized sensors for analysis on-site. We start with a short introduction to CRISPR/Cas systems and the different effector proteins and continue with reviewing the recent developments of integrating CRISPR sensing in miniaturized sensors for point-of-care applications. Finally, we discuss the challenges of point-of-care CRISPR sensing and describe future research perspectives.