Beschreibung
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) systems confer adaptive immunity against viruses and other genetic elements in the majority of Archaea and many Bacteria. Small CRISPR RNAs (crRNAs) are transcribed and processed from genomic CRISPR loci and incorporated into specific CRISPR effector complexes which further degrade foreign nucleic acids complementary to the crRNA. A type III-B CRISPR/CMR system in the hyperthermophilic archaeon Sulfolobus solfataricus was shown to target invader RNA in vitro (Zhang et al., 2012) and in vivo (Zebec et al., 2014). Building on those observations, we designed an artificial miniCRISPR locus expressing crRNAs that match a host mRNA at a defined position to trigger targeted gene silencing. Indeed, 50% of ?-galactosidase activity was knocked down using this artificial miniCRISPR (Zebec et al., 2014). Recent multiplexing studies indicate a gradual increase of silencing (up to 95%) when the host was challenged with an increased number of crRNAs expressed from one miniCRISPR targeting the same mRNA at a single or different positions (Zebec and Zink et al., in review). Thus silencing via the CRISPR/CMR system represents a promising way to elucidate the role of essential genes in hyperthermophiles, for which no gene silencing tools have yet been available.Zeitraum | 1 Aug. 2016 |
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Ereignistitel | Molecular Biology of Archaea 5 (MBoA5) |
Veranstaltungstyp | Konferenz |
Ort | London, Großbritannien / Vereinigtes KönigreichAuf Karte anzeigen |