US 12,416,099 B2
Methods of optimizing gene expression in plant
Dror Shalitin, Raanana (IL); Noam Grimberg, Kibutz Lehavot Haviva (IL); Daniel Bechar, Kibbutz Givat Haim Meuhad (IL); and Tal Cohen, Herzelyia (IL)
Assigned to PLANTARC BIO LTD., Raanana (IL)
Filed by PLANTARC BIO LTD., Raanana (IL)
Filed on Sep. 23, 2024, as Appl. No. 18/892,893.
Application 18/892,893 is a continuation of application No. PCT/IL2023/050380, filed on Apr. 9, 2023.
Claims priority of application No. 292199 (IL), filed on Apr. 12, 2022.
Prior Publication US 2025/0059531 A1, Feb. 20, 2025
Int. Cl. C40B 30/06 (2006.01)
CPC C40B 30/06 (2013.01) 12 Claims
OG exemplary drawing
 
1. A method for in-planta high throughput evaluation of endogenous genetic regulatory elements of a crop plant, the method comprising the steps of:
a. obtaining a nucleic acid of a native promoter regulatory element of a trait-related gene, coupling it to a coding sequence of a reporter gene having a native function in the crop plant; wherein the regulatory element is of the crop plant;
b. introducing one or more random genetic changes in each of a plurality of copies of the native regulatory element, thereby obtaining a genetic library comprising a large collection of altered regulatory elements;
c. introducing the genetic library into model plants, utilizing high throughput stable transformation, such that on average each model plant receives a single altered regulatory element;
d. screening the transformed model plants, wherein the screening comprises selecting a model plant having a desired change in phenotype thereof, said change is correlated to expression level of the reporter gene; and
e. identifying the one or more genetic changes in the altered regulatory element of the selected model plant.