use of at least one rna guide molecule and cas protein, method of binding, cleaving, labeling or modifying a double-stranded target polynucleotide, non-human transformed cell, and nucleoprotein complex

摘要

thermostable cas9 nucleases. The present invention relates to the field of genetic engineering and, more particularly, to nucleic acid editing and genome modification. The present invention provides an isolated cas protein or polypeptide fragment thereof having an amino acid sequence of sequence id: 1 or a sequence of at least 77% identity to it. the cas protein or polypeptide is capable of binding, cleaving, labeling or modifying a double-stranded target polynucleotide at a temperature in the range of 20 ° C and up to and including 100 ° C. The invention further provides isolated nucleic acid molecules encoding said cas9 nucleases, expression vectors and host cells. The invention also provides pam sequences recognized by the cas protein or polypeptide. the cas9 nucleases described herein provide new tools for general genetic engineering, particularly at elevated temperatures, and are of particular value in the genetic manipulation of thermophilic organisms; particularly microorganisms.