Decentralizing Cell-Free RNA Sensing With the Use of Low-Cost Cell Extracts

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dc.contributor.authorArce, Anibal
dc.contributor.authorGuzman Chavez, Fernando
dc.contributor.authorGandini, Chiara
dc.contributor.authorPuig, Juan
dc.contributor.authorMatute, Tamara
dc.contributor.authorHaseloff, Jim
dc.contributor.authorDalchau, Neil
dc.contributor.authorMolloy, Jenny
dc.contributor.authorPardee, Keith
dc.contributor.authorFederici, Fernan
dc.date.accessioned2025-01-20T22:08:52Z
dc.date.available2025-01-20T22:08:52Z
dc.date.issued2021
dc.description.abstractCell-free gene expression systems have emerged as a promising platform for field-deployed biosensing and diagnostics. When combined with programmable toehold switch-based RNA sensors, these systems can be used to detect arbitrary RNAs and freeze-dried for room temperature transport to the point-of-need. These sensors, however, have been mainly implemented using reconstituted PURE cell-free protein expression systems that are difficult to source in the Global South due to their high commercial cost and cold-chain shipping requirements. Based on preliminary demonstrations of toehold sensors working on lysates, we describe the fast prototyping of RNA toehold switch-based sensors that can be produced locally and reduce the cost of sensors by two orders of magnitude. We demonstrate that these in-house cell lysates provide sensor performance comparable to commercial PURE cell-free systems. We further optimize these lysates with a CRISPRi strategy to enhance the stability of linear DNAs by knocking-down genes responsible for linear DNA degradation. This enables the direct use of PCR products for fast screening of new designs. As a proof-of-concept, we develop novel toehold sensors for the plant pathogen Potato Virus Y (PVY), which dramatically reduces the yield of this important staple crop. The local implementation of low-cost cell-free toehold sensors could enable biosensing capacity at the regional level and lead to more decentralized models for global surveillance of infectious disease.
dc.fuente.origenWOS
dc.identifier.doi10.3389/fbioe.2021.727584
dc.identifier.issn2296-4185
dc.identifier.urihttps://doi.org/10.3389/fbioe.2021.727584
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/94311
dc.identifier.wosidWOS:000693915900001
dc.language.isoen
dc.revistaFrontiers in bioengineering and biotechnology
dc.rightsacceso restringido
dc.subjectcell-free
dc.subjecttoehold-sensor
dc.subjectlow-cost
dc.subjectdiagnostics
dc.subjectdecentralization
dc.subject.ods03 Good Health and Well-being
dc.subject.odspa03 Salud y bienestar
dc.titleDecentralizing Cell-Free RNA Sensing With the Use of Low-Cost Cell Extracts
dc.typeartículo
dc.volumen9
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
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