US 12,378,566 B2
Plant genome modification using guide RNA/Cas endonuclease systems and methods of use
Andrew Mark Cigan, Madison, WI (US); Saverio Carl Falco, Wilmington, DE (US); Huirong Gao, Johnston, IA (US); Zhongsen Li, Hockessin, DE (US); Zhan-Bin Liu, Clive, IA (US); L. Aleksander Lyznik, Johnston, IA (US); Jinrui Shi, Johnston, IA (US); Sergei Svitashev, Johnston, IA (US); and Joshua K. Young, Johnston, IA (US)
Assigned to PIONEER HI-BRED INTERNATIONAL, INC., Johnston, IA (US); and E.I. DU PONT DE NEMOURS AND COMPANY, Wilmington, DE (US)
Filed by E. I. DU PONT DE NEMOURS AND COMPANY, Wilmington, DE (US); and PIONEER HI-BRED INTERNATIONAL, INC., Johnston, IA (US)
Filed on Nov. 15, 2022, as Appl. No. 18/055,614.
Application 18/055,614 is a continuation of application No. 14/913,614, abandoned, previously published as PCT/US2014/051778, filed on Aug. 20, 2014.
Claims priority of provisional application 62/023,239, filed on Jul. 11, 2014.
Claims priority of provisional application 61/953,090, filed on Mar. 14, 2014.
Claims priority of provisional application 61/937,045, filed on Feb. 7, 2014.
Claims priority of provisional application 61/882,532, filed on Sep. 25, 2013.
Claims priority of provisional application 61/868,706, filed on Aug. 22, 2013.
Prior Publication US 2023/0323374 A1, Oct. 12, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/82 (2006.01); C12N 15/00 (2006.01); C12N 15/63 (2006.01); A01H 1/00 (2006.01)
CPC C12N 15/8213 (2013.01) [C12N 15/00 (2013.01); C12N 15/63 (2013.01); C12N 15/8205 (2013.01); C12N 15/8207 (2013.01); C12N 15/8216 (2013.01); C12N 15/8247 (2013.01); C12N 15/8262 (2013.01); A01H 1/00 (2013.01)] 18 Claims
 
1. A method for modifying a target site in the genome of a plant cell, the method comprising:
transforming at least one plant cell with a guide polynucleotide, a polynucleotide sequence encoding a Cas endonuclease, and an expression cassette encoding at least one developmental gene polypeptide, wherein the at least one developmental gene polypeptide comprises a Babyboom (BBM), an Ovule Development Protein 2 (ODP2), and/or a Wuschel (WUS) polypeptide, wherein the guide polynucleotide and the Cas endonuclease form a guide polynucleotide-Cas endonuclease complex that introduces a double-strand break at the target site in the plant cell, and wherein the at least one developmental gene polypeptide increases the frequency of modification of the target site by the guide polynucleotide-Cas endonuclease complex in the plant cell relative to a control plant cell comprising a guide polynucleotide-Cas endonuclease complex and not comprising an expression cassette encoding a developmental gene polypeptide; and
regenerating a plantlet or a plant from the plant cell, the plantlet or plant comprising a modified target site, wherein the plantlet or the plant does not contain the Cas endonuclease or the expression cassette encoding the at least one developmental gene polypeptide.