The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.
The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.
Patent No.:
Date of Patent:
Aug. 27, 2024
Filed:
Dec. 14, 2017
Wageningen Universiteit, Wageningen, NL;
Stichting Voor DE Technische Wetenschappen, Utrecht, NL;
John Van Der Oost, Renkum, NL;
Richard Van Kranenburg, Gorinchem, NL;
Elleke Fenna Bosma, Denmark, NL;
Ioannis Mougiakos, Wageningen, NL;
Prarthana Mohanraju, Wageningen, NL;
Wageningen Universiteit, Wageningen, NL;
Stichting Voor De Technische Wetenschappen, Utrecht, NL;
Abstract
A polynucleotide encoding a ThermoCas9 protein fromand a constitutive promoter are used to engineer eukaryotic cells, e.g. fungi, yeast or algae, so that the ThermoCas9 endonuclease is integrated and expressed from the genome of the cell. Then, a second expression plasmid is used to transfect these ThermoCas9 expressing cells, the second plasmid containing an inducible promoter and a polynucleotide encoding a guide RNA. The guide RNA combines with the ThermoCas9 to provide the targeted endonuclease activity to cleave the cell DNA at a desired locus or gene of interest. A repair-oligo is also provided to the cell whereby following DNA cleavage, homologous recombination takes place in the cell with the repair-oligo so that either a deletion or substitution of nucleotides in the locus or gene of interest is achieved. Expression vectors and methods of using the vectors to achieve ThermoCas9 mediated gene editing are described whereby higher temperatures, e.g. greater than 30° C., are used.