Master's degree project in genetic engineering
Available earliest April 1st 2018
Alyona Minina, PhD: alena.minina (@) slu.se
Anna Åsman, PhD: anna.asman (@) slu.se
Most of the current plant molecular biology studies still rely on the use of crude genetic engineering tools that dramatically limit the capacity of our research. The recent advances in the use of CRISPR-Cas9 system for plants give very promising results that still require some significant modifications.
In this project we aim to optimize the CRISPR-Cas9 for precise knock-in modification of Arabidopsis genes and use the new tool to make reporter lines for detection of plant autophagy-related (ATG) genes activity.
This project, in general, will open up a broad range of new possibilities for investigating plant gene function and in particular, will make a significant contribution to our understanding of ATG-genes regulation.
Establish proof of concept constructs for knock-in modification of Arabidopsis thaliana genes in protoplasts
Create a set of constructs for knock-in modification of genes important for regulation of autophagy in Arabidopsis thaliana
Participate in establishing transgenic lines by knocking in green fluorescent protein and luciferases into Arabidopsis genome
You will acquire skills in
Use of CRISPR-Cas9 in plants
Advanced DNA and protein molecular biology methods
Advanced confocal microscopy
Handling typical plant model organisms: Arabidopsis thaliana plants and tobacco cell cultures
An example of CRISPR-Cas9 driven knock-in. A stop codon of a gene in Arabidopsis genome is replaced with a DNA sequence encoding theGreen Fluorescent Protein (GFP). The resulting plant expresses a GFP fusion of the endogenous protein.