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Substrate Generation for Endonucleases of CRISPR/Cas Systems
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September 08, 2012
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CRISPR/Cas systems mediate adaptive immunity in Bacteria and Archaea. Many Cas proteins are proposed to act as endoribonucleases acting on crRNA precursors of varying length. Here we illustrate three different approaches to generate pre-crRNA substrates for the biochemical analysis of Cas endonuclease activity.
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Substrate GenerationEndonucleasesCRISPR/Cas SystemsVirusesProkaryotic HostsBacterial EvolutionArchaeal LifeRestriction ModificationAbortive InfectionAdaptive Immune SystemsCRISPR SequencesCas GenesCas ProteinsSpacer ElementCRISPR ClusterPrecursor-crRNACas6 FamilyCrRNAsViral DNA UptakeCascade ComplexBase ComplementarityNuclease Cas3Invader DNA Destruction
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Zoephel, J., Dwarakanath, S., Richter, H., Plagens, A., Randau, L. Substrate Generation for Endonucleases of CRISPR/Cas Systems. J. Vis. Exp. (67), e4277, doi:10.3791/4277 (2012).
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