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Platform for precise allele conversion using synthetic endonucleases (PRECISE)
Project
Project code: 031B0547
Contract period: 01.06.2018
- 31.05.2020
Budget: 316,338 Euro
Purpose of research: Basic research
The overall goal of PRECISE is to provide a pipeline for precise allele replacement in cereals which enable an improved breeding process by directly modifying weak alleles in elite breeding material without the need of time-consuming and laborious back-crossing. Therefore in a direct comparison commercial available SpCas9 and FnCpf1 proteins will be used for site-directed mutagenesis in barley. After method establishment in leaves immature embryos of barley will be transformed to generate respective mutant plants. Optimal conditions identified to target one gene will be transferred for targeting two or more genes at the same time. To promote homology-directed repair key genes of the non-homologous endjoining pathway (NHEJ) will be repressed by RNA interference. We will apply commercially available siRNAs specific for the before mentioned genes to the immature embryos prior to biolistic transformation with the ribonucleoprotein complex (gRNA and endonuclease) using the same targets as above. Alternatively we will apply pharmaceutical inhibitors. Different concentrations and incubation times will be compared to find the optimal condition. To tackle the problem of providing optimal repair template several template characteristics will be optimized. Starting with single targets at the end multiplexing of several targets at the same time shall be achieved.
Section overview
Subjects
- Plant Breeding
- Process engineering
Framework programme
Funding programme
Excutive institution
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)
