Project properties
Title CRISPR-Cas in plant development
Group Molecular Biology, Laboratory of
Project type thesis
Credits 18-39
Supervisor(s) Dr. Ir. R Heidstra
Examiner(s) prof dr B. Scheres (PDB/MOB)
Contact info renze.heidstra@wur.nl
Begin date 2017/09/01
End date 2024/10/01
Description Recent advances with the RNA-mediated CRISPR-Cas systems have dramatically transformed our ability to specifically modify intact genomes of diverse cells and organisms. The CRISPR–Cas system has proven to be an efficient, simple, and robust gene-targeting technology with enormous potential for applications across basic science, agricultural and biotechnology.

We have standardized the CRISPR-Cas technology for efficient use in Arabidopsis with the purpose to create in vivo specific mutations and deletions in the genome for genes involved in plant development. We target genes involved in stem cell biology, lateral root formation and cell division/differentiation. Particularly useful is the ability to perform multiplex CRISPR-Cas to either generate multiple mutations or to delete genes from the genome.

You generate the different modules that are used to generate CRISPR-Cas construct. For this purpose you will get familiar with the modular Golden Gate cloning technology which presents a directional method to construct vectors carrying multiple inserts. Finally, you will perform genome editing and analyse the mutations/deletions created, both molecularly and phenotypically.
Used skills Standard recombinant DNA technology such as DNA isolation, PCR and (Golden Gate) cloning.
Handling and transformation of model organisms; E.coli, agrobacterium, arabidopsis.
Gene expression analysis by way of quantitative RT-PCR.
Analysis of gene knock-outs by way of PCR-genotyping and microscopy-phenotyping.
Requirements For BSc thesis: MOB20306
For MSc thesis: MOB20306 and and MOB30306 or MOB31303 or MOB30806 or PHP30806 or equivalent