	US 11,970,710 B2
	Genome engineering with Type I CRISPR systems in eukaryotic cells
Charles A. Gersbach, Durham, NC (US); Adrian Pickar Oliver, Durham, NC (US); and Chase Beisel, Raleigh, NC (US)
Assigned to Duke University, Durham, NC (US); and North Carolina State University, Raleigh, NC (US)
Appl. No. 15/766,912
Filed by Duke University, Durham, NC (US); and North Carolina State University, Raleigh, NC (US)
PCT Filed Oct. 13, 2016, PCT No. PCT/US2016/056925 § 371(c)(1), (2) Date Apr. 9, 2018, PCT Pub. No. WO2017/066497, PCT Pub. Date Apr. 20, 2017.
Claims priority of provisional application 62/382,240, filed on Aug. 31, 2016.
Claims priority of provisional application 62/240,716, filed on Oct. 13, 2015.
Claims priority of provisional application 62/240,743, filed on Oct. 13, 2015.
Prior Publication US 2018/0334688 A1, Nov. 22, 2018
Int. Cl. C12N 15/90 (2006.01); C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01)

CPC C12N 15/907 (2013.01) [C12N 9/22 (2013.01); C12N 15/102 (2013.01); C12N 15/11 (2013.01); C07K 2319/00 (2013.01); C07K 2319/09 (2013.01); C07K 2319/21 (2013.01); C07K 2319/41 (2013.01); C07K 2319/43 (2013.01); C07K 2319/80 (2013.01); C12N 2310/20 (2017.05); C12N 2740/15043 (2013.01); C12N 2740/16043 (2013.01); C12N 2800/22 (2013.01); C12N 2830/001 (2013.01)]

17 Claims

1. A composition for genome engineering a target gene in a eukaryotic cell, the composition comprising:

a. a polynucleotide sequence encoding:

i. a Cascade complex comprising a Cse1 polypeptide, a Cse2 polypeptide, a Cas7 polypeptide, a Cas5 polypeptide, and a Cas6e polypeptide,

wherein the polynucleotide sequence encoding the Cse1 polypeptide comprises the polynucleotide sequence of SEQ ID NO: 23, the polynucleotide sequence encoding the Cse2 polypeptide comprises the polynucleotide sequence of SEQ ID NO: 24, the polynucleotide sequence encoding the Cas7 polypeptide comprises the polynucleotide sequence of SEQ ID NO: 25, the polynucleotide sequence encoding the Cas5 polypeptide comprises the polynucleotide sequence of SEQ ID NO: 26, and the polynucleotide sequence encoding the Cas6e polypeptide comprises the polynucleotide sequence of SEQ ID NO: 27; and

ii. a polypeptide domain, wherein the polypeptide domain comprises transcription activation activity, transcription repression activity, transcription release factor activity, histone modification activity, nuclease activity, nucleic acid association activity, methylase activity, or demethylase activity; and

b. a pharmaceutically acceptable carrier.