US 12,460,219 B2
Methods and systems for targeted modification of plant genes via meristem modulation and plants having engineered meristem modifications and other mutations
Brian G. Ayre, Denton, TX (US); and Roisin Carrie McGarry, Denton, TX (US)
Assigned to UNIVERSITY OF NORTH TEXAS, Denton, TX (US)
Filed by UNIVERSITY OF NORTH TEXAS, Denton, TX (US)
Filed on May 20, 2022, as Appl. No. 17/749,560.
Claims priority of provisional application 63/191,072, filed on May 20, 2021.
Prior Publication US 2022/0372503 A1, Nov. 24, 2022
Int. Cl. C12N 15/82 (2006.01)
CPC C12N 15/8218 (2013.01) [C12N 15/8262 (2013.01)] 11 Claims
 
1. A method of producing a genetically modified plant, the method comprising:
silencing a meristem maintenance gene in the plant to induce a phase change in the plant meristem from an indeterminate to a determinate state; and
genetically manipulating a second gene in the plant;
wherein genetic manipulation of the second gene in combination with silencing the meristem maintenance gene results in a detectable new phenotype in the genetically modified plant;
wherein the new phenotype is distinguishable from a phenotype resulting from either silencing the meristem maintenance gene only or mutation of the meristem gene only;
wherein the plant is a cotton plant;
wherein the meristem maintenance gene is SELF-PRUNING (GhSP) from cotton having at least 99% identity to SEQ ID NO: 1;
wherein the second gene is a cotton gene selected from the group consisting of: CLAVATA-WUSCHEL circuit, CLAVATA1, CLAVATA2, CLAVATA3, AGAMOUS (GhAG) from cotton, FASCIATED INFLORESCENCE (FIN), PERIANTHIA, AGAMOUS, and orthologs of the foregoing;
wherein the silencing the meristem maintenance gene is by virus induced gene silencing (VIGS) using a suitable region of SEQ ID NO:1;
wherein the silencing a meristem maintenance gene and the second gene are co-delivered; and
wherein the genetically manipulating a second gene is by:
(a) co-silencing the second gene with the meristem maintenance gene;
(b) editing the second gene with delivery of RNA molecules and/or RNA-based signals;
(c) modifying the second gene with a CRISPR/Cas gene editing system comprising a Cas nuclease and one or more CRISPR RNA guide sequences targeting the second gene; or
(d) virus induced gene silencing (VIGS).