Sirvi Autor "Yeh, Chung-Yueh, juhendaja" järgi

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Using CRISPR/cas9 to verify the cause of conditional sterility in Arabidopsis thaliana harbouring a mutated MPK12
(Tartu Ülikool, 2021) Mäll, Karl; Wang, Yuh-Shuh, juhendaja; Yeh, Chung-Yueh, juhendaja; Tartu Ülikool. Loodus- ja täppisteaduste valdkond; Tartu Ülikool. Molekulaar- ja rakubioloogia instituut
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.