Nucleic acid aptamers and enzymes as sensors

aptamer 核酸识体,酶与生物传感器,英文综述。

Nucleic acid aptamers and enzymes as sensors

Nucleic acid aptamers and enzymes as sensors

Nucleicacidaptamersandenzymesassensors

NaveenKNavaniandYingfuLi

ThefunctionofnucleicacidshasbeenanendlesssourceofdiscoveryandinventionthathasdrasticallyenhancedourappreciationofDNAandRNAasmultifacetedpolymers.Itisnowwidelyknownthatnucleicacidscanactasenzymes(deoxyribozymesandribozymes)andasreceptors(aptamers),andthatthesefunctionalnucleicacids(FNAs)caneitherbefoundinnatureorisolatedfrompoolsofrandomnucleicacids.Theavailabilityofmanynaturalandarti cialFNAshasopenedanewhorizonforthedevelopmentof‘smart’moleculesforavarietyofchemicalandbiologicalapplications.ThisreviewprovidesasnapshotofrecentprogressintheapplicationofFNAsasnovelsensorsforbiomoleculardetection,drugdiscoveryandnanotechnology.

Addresses

DepartmentofBiochemistryandBiomedicalSciencesand

DepartmentofChemistry,McMasterUniversity,1200MainStreetWest,Hamilton,Ontario,L8N3Z5,Canada

Correspondingauthor:Li,Yingfu(liying@mcmaster.ca)

tonucleicacidaptamersandenzymescollectivelyas‘functionalnucleicacids’orsimply‘FNAs’.Althoughabroadde nitionofFNAswouldalsoincludehybridiza-tion-basedprobessuchasstandardmolecularbeaconsandantisenseoligonucleotides,wehavechosentofocusonaptamersandnucleicacidenzymesinthestrictsense.

Functionalnucleicacidsassensors:generalconsiderations

FNAsareattractiveoptionsforsensingapplications.TheycanbegeneratedbySELEXtobinddiversetargetsincludingthoseforwhichantibodiesaredif culttoobtain(suchastoxins).Moreover,invitroselectioncanbeusedtoevolveFNAsunderanypre-de nedconditions,mak-ingitpossibletodesignsensorsfortasksthatcannotbemetwithproteinreceptors.Nucleicacidscanalsobeeasilyimmobilizedtoprovideacustom-madesurfaceforspeci capplications.Theyhavetheaddedadvantageofbeingabletorefoldtotheirnativeconformationfollowingoneroundofsensing,andthereforerepresentreusabledevices.ThechangeinshapeofFNAsuponbindingtotheirtargetcanbeconvenientlycoupledwithvarioussignalingmechanismsforeasymonitoringofmolecularrecognitionevents.

Abiosensorisgenerallyde nedasananalyticaldeviceconsistingofabiologicallyrelevantmolecularrecognitionelement(MRE)integratedtoasignaltransductionele-ment(Figure1a).Inthisreview,wewill rstexaminetheuseofFNAsforeasydetectionofproteins,nucleicacidsandmetabolites,followedbyadiscussiononsomeofthelatestactivitiesinvolvingFNAsastoolsindrugdiscoveryandnanotechnology.

CurrentOpinioninChemicalBiology2006,10:272–281Thisreviewcomesfromathemedissueon

Combinatorialchemistryandmoleculardiversity

EditedbyScottKSilvermanandPaulJHergenrotherAvailableonline4thMay2006

1367-5931/$–seefrontmatter

#2006ElsevierLtd.Allrightsreserved.DOI10.1016/j.cbpa.2006.04.003

Introduction

Ourappreciationofthefunctionalversatilityofnucleicacidshasgrownoverthepasttwodecades.AquickperusaloftheliteraturerevealsthatmanyfunctionalRNAs,includingribozymes,microRNAsandribos-witches,existinnature[1–3].Furthermore,manyman-madeaptamers,ribozymes,deoxyribozymesandallos-tericnucleicacidenzymeshavebeengeneratedvia‘invitroselection’or‘SELEX’(systemicevolutionofligandsbyexponentialenrichment)techniques[4,5].Thereali-zationthatnucleicacidaptamersandenzymesarehighlyusefulmoleculartoolshasmadethestudyoffunctionalnucleicacidsaveryimportantpartofchemicalbiology.The exibilityofferedbynucleicacids(stability,easeofimmobilizationandsusceptibilitytovariouschemicalmodi cationsandlabeling)hasmotivatedmolecularscientistsworldwidetoseekinnovativeapplicationsforthesespecies.Thisreviewdiscussessomenewadvancesthatharnessthepotentialofnucleicacidaptamersandenzymesassensormolecules.Forsimplicity,wewillrefer

CurrentOpinioninChemicalBiology2006,10:272–281

Functionalnucleicacidsassensorsforproteindetection

Manyprotein-bindingaptamershavebeenisolated,anumberofwhichhavebeenexploitedforproof-of-con-ceptbiosensingapplicationsusingvarioussignaltrans-ductionmechanisms.

Opticalsensing

Opticaldetectionby uorescencespectroscopyisapop-ularmethod,duelargelytotheeasewithwhichFNAscanbe uorescentlylabelled,theavailabilityofmanydiffer-ent uorophoresandquenchers,andtheinherentcap-abilityforreal-timemultiplexdetection.Severalstrategieshavebeendescribedforconvertinganexistingaptamerintoa uorescentprobe(recentlyreviewedin[6]).Twofrequentlyadoptedmethodsarethemolecularbeaconapproach(Figure1b)[7]andtheduplex-to-com-plexswitchingapproach(Figure1c)[8].Theformer

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