Abstract: The present invention describes a novel bacterial nuclease of the CRISPR-Cas9 system from the bacterium P. pneumotropica, 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 modifying the genomic DNA sequence in the cell of a unicellular organism or a multicellular organism. Thus, the versatility of the available CRISPR-Cas9 systems is increased, which fact will enable the use of various variants of Cas9 nucleases for cutting genomic or plasmid DNA in various organisms, in a larger number of specific sites and/or under various conditions.

Abstract: The present invention relates to the field of biotechnology, molecular biology and medicine, in particular to nuclease enzyme and use thereof. More specifically, the present invention relates to PaCas9 nuclease enzyme. The invention also relates to a nucleic acid encoding said nuclease, a genetic construct, an expression vector, a delivery vector, which comprise said nucleic acid, a liposome comprising said nuclease or nucleic acid encoding said nuclease, a method for producing a nuclease, methods for delivery, and a host cell comprising a nucleic acid encoding said nuclease.

Abstract: The present invention describes a novel bacterial nuclease of the CRISPR-Cas9 system from the bacterium Defluviimonas sp. 20V17, 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 specific conditions.

Abstract: 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.