	US 12,071,639 B2
	Thermostable Cas9 nucleases
John Van Der Oost, Renkum (NL); Richard Van Kranenburg, Gorinchem (NL); Elleke Fenna Bosma, Denmark (NL); Ioannis Mougiakos, Wageningen (NL); and Prarthana Mohanraju, Wageningen (NL)
Assigned to Wageningen Universiteit, Wageningen (NL); and Stichting Voor De Technische Wetenschappen, Utrecht (NL)
Appl. No. 16/469,691
Filed by Wageningen Universiteit, Wageningen (NL); and Stichting Voor De Technische Wetenschappen, Utrecht (NL)
PCT Filed Dec. 14, 2017, PCT No. PCT/EP2017/082870 § 371(c)(1), (2) Date Jun. 14, 2019, PCT Pub. No. WO2018/109101, PCT Pub. Date Jun. 21, 2018.
Claims priority of application No. PCT/EP2016/081077 (WO), filed on Dec. 14, 2016; and application No. PCT/EP2017/070796 (WO), filed on Aug. 16, 2017.
Prior Publication US 2019/0360002 A1, Nov. 28, 2019
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01); C12N 15/113 (2010.01); C12N 15/90 (2006.01)

CPC C12N 9/22 (2013.01) [C12N 15/102 (2013.01); C12N 15/11 (2013.01); C12N 15/113 (2013.01); C12N 15/902 (2013.01); C12N 15/907 (2013.01); C12N 2310/20 (2017.05); C12N 2330/51 (2013.01)]

17 Claims

1. A polynucleotide expression vector for modifying the genetic material at a target locus of a host organism which comprises the expression vector, the vector comprising:

a) a polynucleotide sequence encoding a ThermoCas9 nuclease, wherein the ThermoCas9 nuclease comprises the amino acid sequence of SEQ ID NO: 1 or a sequence of at least 89% identity therewith, or an active fragment thereof;

b) a polynucleotide sequence encoding a guide RNA, wherein the guide RNA has a nucleic acid sequence which recognizes a target nucleic acid sequence comprised in the target locus; and

c) a first promoter oriented with respect to polynucleotide sequences of (a) and (b) so as to drive expression thereof in the organism, wherein said first promoter is an inducible promoter.