	US 12,473,564 B2
	Methods and compositions for generating dominant alleles using genome editing
Edward James Cargill, Chesterfield, MO (US); Douglas Michael Eudy, Maxwell, IA (US); Andrei Y. Kouranov, Chesterfield, MO (US); Richard Joseph Lawrence, Kirkwood, MO (US); Thomas L. Slewinski, Chesterfield, MO (US); Randy Shultz, Davis, CA (US); PokChun Jennifer To, Durham, NC (US); Samuel Sukhwan Yang, Ballwin, MO (US); and Yuanji Zhang, Chesterfield, MO (US)
Assigned to MONSANTO TECHNOLOGY LLC, St. Louis, MO (US)
Filed by MONSANTO TECHNOLOGY LLC, St. Louis, MO (US)
Filed on May 28, 2020, as Appl. No. 16/886,683.
Claims priority of provisional application 62/854,142, filed on May 29, 2019.
Claims priority of provisional application 62/886,726, filed on Aug. 14, 2019.
Claims priority of provisional application 62/886,732, filed on Aug. 14, 2019.
Prior Publication US 2020/0377900 A1, Dec. 3, 2020
Int. Cl. C12N 15/82 (2006.01)

CPC C12N 15/8218 (2013.01) [C12N 2310/11 (2013.01); C12N 2310/127 (2013.01); C12N 2310/141 (2013.01)]

6 Claims

1. A method of generating a dominant negative allele of a gene in a cell comprising inverting a portion of the gene using a targeted editing technique, wherein the inversion results in the production of an antisense RNA transcript under the control of the native promoter of the gene, wherein the antisense RNA transcript is capable of triggering suppression of an unmodified allele of the gene, wherein the dominant negative allele is not exogenously introduced, wherein the gene is a GA20 oxidase gene, and wherein the GA20 oxidase gene is encoded by an mRNA that encodes a protein having at least 95% sequence identity to a sequence selected from the group consisting of SEQ ID NO: 30 and SEQ ID NO: 35.