US 12,297,439 B2
Compositions and methods for driving T1 event diversity
Timothy Kelliher, Research Triangle Park, NC (US); Jiang Li, Beijing (CN); Julie Leonard Green, Woodland, CA (US); Zhongying Chen, Research Triangle Park, NC (US); Wan Shi, Beijing (CN); and Guozhu Tang, Research Triangle Park, NC (US)
Assigned to Syngenta Crop Protection AG, Basel (CH)
Appl. No. 17/429,125
Filed by Syngenta Crop Protection AG, Basel (CH)
PCT Filed Feb. 24, 2020, PCT No. PCT/US2020/019499
§ 371(c)(1), (2) Date Aug. 6, 2021,
PCT Pub. No. WO2020/176412, PCT Pub. Date Sep. 3, 2020.
Claims priority of application No. PCT/CN2019/076062 (WO), filed on Feb. 25, 2019.
Prior Publication US 2022/0145312 A1, May 12, 2022
Int. Cl. C12N 15/82 (2006.01)
CPC C12N 15/8213 (2013.01) [C12N 15/8231 (2013.01); C12N 15/8233 (2013.01)] 20 Claims
 
1. A method for producing a plurality of unique edits in a plant's T1 seed, the method comprising:
a) transforming at least one expression cassette into a plant cell or a plant tissue, wherein the at least one expression cassette comprises
a nucleic acid that encodes a DNA modification enzyme,
a nucleic acid that encodes at least one guide RNA (gRNA), and
a floral mosaic (FMOS) regulatory sequence,
wherein the FMOS regulatory sequence
(i) mediates expression of the DNA modification enzyme in at least one of a floral primordia cell and a floral reproductive organ, and
(ii) mediates a plurality of edits in the at least one of the floral primordia and the floral reproductive organ; and
b) regenerating the plant cell or plant tissue into a T0 plant having a plurality of T1 seed, wherein the T1 seed contain a plurality of unique edits;
wherein the FMOS regulatory sequence comprises an FMOS promotor selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 20, SEQ ID NO: 26, SEQ ID NO: 29, SEQ ID NO: 35, SEQ ID NO: 47, SEQ ID NO: 50, SEQ ID NO: 53, SEQ ID NO: 56, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO: 71, SEQ ID NO: 74, SEQ ID NO: 77, SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 86, SEQ ID NO: 514, SEQ ID NO: 518, SEQ ID NO: 614, SEQ ID NO: 615, SEQ ID NO: 616, SEQ ID NO: 617, SEQ ID NO: 618, SEQ ID NO: 619, SEQ ID NO: 620, SEQ ID NO: 621, SEQ ID NO: 622, SEQ ID NO: 698, SEQ ID NO: 700, SEQ ID NO: 702, SEQ ID NO: 704, SEQ ID NO: 706, SEQ ID NO: 708, SEQ ID NO: 710, SEQ ID NO: 717, SEQ ID NO: 719, SEQ ID NO: 721,
and SEQ ID NO: 723 or a sequence with at least 95% sequence identity thereto.