Using CRISPR/cas9 to verify the cause of conditional sterility in Arabidopsis thaliana harbouring a mutated MPK12
Date
2021
Authors
Journal Title
Journal ISSN
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Publisher
Tartu Ülikool
Abstract
MPK12 is a gene that exists in a number of plants and has many functions, however the
development of seeds is not among them. Arabidopsis thaliana plants with a specific mutational
variant G187R of MPK12 have what is referred to as conditional sterility and are unable to
produce seeds at the usual growth temperatures. To confirm whether the mutation MPK12
(G187R) is responsible for the sterility, genetic modifications and DNA sequencing are used.
First, a genotype analysis by DNA sequencing was compared to the flowering phenotype to
determine whether the G187R mutation was linked to the sterility. Additionally, a CRISPR/cas9
approach was used to knockout the mutated MPK12. If the sterility phenotype is caused by the
G187R mutation knock-out of that mutated gene should reverse it. In the paper two constructs
with differing methods of replication, are designed and constructed to deliver the CRISPR/cas9
reagents to Arabidopsis via Agrobacterium-mediated transformation.
Description
Keywords
MPK12, sterility, CRISPR/cas9, Arabidopsis thaliana