{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,3]],"date-time":"2026-05-03T17:05:47Z","timestamp":1777827947851,"version":"3.51.4"},"reference-count":52,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2011,3,8]],"date-time":"2011-03-08T00:00:00Z","timestamp":1299542400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Microbiology"],"published-print":{"date-parts":[[2011,4]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>Clustered regularly interspaced short palindromic repeats (CRISPR)\/Cas systems are found widespread in bacterial and archaeal genomes and exhibit considerable diversity. However, closer insights into the action of most of the CRISPR modules have remained elusive in particular in Archaea as a result of the lack of suitable <jats:italic>in vivo<\/jats:italic> test systems. Here we demonstrate CRISPR\/Cas\u2010based immune defence in the hyperthermophilic archaeon <jats:italic>Sulfolobus solfataricus<\/jats:italic>. Recombinant variants of the SSV1 virus containing a gene of the conjugative plasmid pNOB8 that represents a target for a corresponding CRISPR spacer in the chromosome were tested in transfection experiments. Almost 100% immunity against the recombinant virus was observed when the chromosomal CRISPR spacer matched perfectly to the protospacer. Different from bacterial systems immunity was still detected, albeit at decreased levels, when mutations distinguished target and spacer. CRISPR\/Cas targeting was independent of the transcription of the target gene. Furthermore, a mini\u2010CRISPR locus introduced on the viral DNA with spacers targeting the (non\u2010essential) chromosomal beta\u2010galactosidase gene was unstable in host cells and triggered recombination with the indigenous CRISPR locus. Our experiments demonstrate <jats:italic>in vivo<\/jats:italic> activity of CRISPR\/Cas in archaea for the first time and suggest that \u2013 unlike the recently demonstrated <jats:italic>in vitro<\/jats:italic> cleavage of RNA in <jats:italic>Pyrococcus<\/jats:italic>\u2013 DNA is targeted in this archaeon.<\/jats:p>","DOI":"10.1111\/j.1365-2958.2011.07586.x","type":"journal-article","created":{"date-parts":[[2011,3,9]],"date-time":"2011-03-09T05:12:42Z","timestamp":1299647562000},"page":"481-491","source":"Crossref","is-referenced-by-count":90,"title":["<i>In vivo<\/i> activity of CRISPR\u2010mediated virus defence in a hyperthermophilic 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