	US 12,241,074 B2
	Genome editing-based crop engineering and production of brachytic plants
Ty J. Barten, Ankeny, IA (US); Edward J. Cargill, Chesterfield, MO (US); Jonathan C. Lamb, Wildwood, MO (US); Bryce Lemke, Nevada, IA (US); Linda A. Rymarquis, High Ridge, MO (US); and Dennis H. Yang, Ballwin, MO (US)
Assigned to MONSANTO TECHNOLOGY LLC, St. Louis, MO (US)
Appl. No. 16/472,140
Filed by Monsanto Technology LLC, St. Louis, MO (US)
PCT Filed Dec. 21, 2017, PCT No. PCT/US2017/067888 § 371(c)(1), (2) Date Jun. 20, 2019, PCT Pub. No. WO2018/119225, PCT Pub. Date Jun. 28, 2018.
Claims priority of provisional application 62/438,370, filed on Dec. 22, 2016.
Prior Publication US 2020/0140874 A1, May 7, 2020
Int. Cl. C12N 15/82 (2006.01); A01H 1/06 (2006.01); A01H 5/10 (2018.01); A01H 6/46 (2018.01); C07K 14/415 (2006.01)

CPC C12N 15/8262 (2013.01) [A01H 1/06 (2013.01); A01H 5/10 (2013.01); A01H 6/4684 (2018.05); C07K 14/415 (2013.01); C12N 15/8213 (2013.01); C12N 15/8261 (2013.01)]

14 Claims

1. A method for producing a semi-dwarf corn plant with reduced ear height and increased stalk diameter, the method comprising:

a. providing to a corn cell a guide RNA that recognizes a target site in a BR2 gene in the corn cell, wherein the guide RNA comprises a target site sequence operably linked to a scaffold sequence comprising SEQ ID NO: 5, wherein the guide RNA acts in association with a CRISPR-based RNA-guided nuclease that creates a strand break at the target site which results in an edited br2 mutant allele, wherein the CRISPR-based RNA-guided nuclease is capable of binding to a protospacer adjacent motif (PAM) for Cas9 or Cpf1, and wherein the target site sequence is selected from the group consisting of SEQ ID NOs: 7-17;

b. generating a corn plant from the corn cell; and

c. selecting the corn plant exhibiting semi-dwarf phenotype, reduced ear height, and increased stalk diameter relative to a non-edited control corn plant.