Effective intracellular delivery of DNA-free CRISPR in plant cells via lipid nanoparticles

Projecttitel: Effective intracellular delivery of DNA-free CRISPR in plant cells via lipid nanoparticles
Projectnummer: LWV19224
Missie: Sleuteltechnologieën
MMIP: ST2 - Biotechnologie en veredeling
Looptijd: 2020 – 2023
Projectleider: Ferdinand Los

This project aims to expand the application of DNA-free CRISPR technology as an R&D tool to commercially relevant crops (relevant knowledge is now mostly confined to tomato). 
Currently, in plants, CRISPR can be applied in in vitro studies and model organisms. 
However, there are very few commercial crops that directly benefit from its use, as many such crops are not amenable to in vitro culture and/or the crossing involved with the generally used transgenic approaches. 

The use of purified CRISPR components can address this issue but is currently hampered by the difficulty of bringing such CRISPR components into the plant cells. Lipid nanoparticles (LNPs) entail the active component of a validated method for transfection of mammalian cells with CRISPR, at very high efficiency and no toxicity. 

In this project, we will develop LNP-based protocols for a broadly applicable platform technology for delivery of CRISPR into plant cells. 
We will validate the technology in different intact tissues of multiple representative commercial crops, making this key technology accessible for a broad range of relevant plant crops.