DNA cutting means based on Cas9 protein from biotechnologically significant bacterium Clostridium cellulolyticum

摘要

The present invention describes a novel bacterial nuclease of the CRISPR-Cas9 system from the bacterium Clostridium celluloliticum, as well as the use thereof to form strictly specific double-strand breaks in a DNA molecule. This nuclease has unusual properties and may be used as a tool for introducing modifications at strictly defined sites in the genomic DNA sequence of unicellular or multicellular organisms. Thus, the versatility of the available CRISPR-Cas9 systems is increased, which will enable the use of Cas9 nucleases from various organisms for cutting genomic or plasmid DNA in a larger number of specific sites and in wider temperature ranges. Further, provided is more facile editing of the genome of the biotechnologically significant bacterium Clostridium celluloliticum.