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CRISPR loci consist of an array of short repeats separated by spacer sequences that match the genome of viruses and plasmids that infect prokaryotes. Transcription of the CRISPR array generates small antisense RNAs that mediate immunity against these invaders. In recent years, there has been a notable increase in the investigation of CRISPR immunity, but studies have been restricted to organisms in which genetic manipulations are possible. Therefore, there is a need for the development of simple genetic tools that facilitate the study of this important pathway. Here we describe the use of CRISPR decoys, plasmids containing a non-transcribed repeat-spacer unit that disrupt CRISPR immunity. We show that decoys abrogate immunity against conjugation in S. epidermidis to levels comparable to a CRISPR deletion mutant. This technique can be used to generate full or spacer-specific CRISPR knockdowns in organisms in which decoy plasmids can be introduced.
Genetic Techniques, Mutagenesis, Conjugation, Genetic, Staphylococcus epidermidis, Clustered Regularly Interspaced Short Palindromic Repeats, RNA, Antisense, Plasmids, Repetitive Sequences, Nucleic Acid
Genetic Techniques, Mutagenesis, Conjugation, Genetic, Staphylococcus epidermidis, Clustered Regularly Interspaced Short Palindromic Repeats, RNA, Antisense, Plasmids, Repetitive Sequences, Nucleic Acid
citations This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically). | 2 | |
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influence This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically). | Average | |
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