Development of versatile genetic tools to study a DYRK family kinase in Caenorhabditis elegans

Suppressor screen;WGS mapping;Genome editing;Biochemistry

Wang, YuemengAndrew, Deborah J. ;

Johns Hopkins University

关键词: Suppressor screen;    WGS mapping;    Genome editing;    Biochemistry;   

Others  :  https://jscholarship.library.jhu.edu/bitstream/handle/1774.2/37917/WANG-DISSERTATION-2015.pdf?sequence=1&isAllowed=y

瑞士|英语

来源: JOHNS HOPKINS DSpace Repository

【 摘 要 】

Screening for suppressors is a common genetic technique to identify mutations in genes that interact with one another. In Caenorhabditis elegans, suppressor screens are commonly used to identify new genes in a pathway or network of interest. I conducted an EMS-induced mutagenesis suppressor screen for suppressors of a temperature-sensitive mutations in the DYRK family kinase MBK-2. The traditional methods for cloning suppressors are tedious and time-consuming. To circumvent these difficulties, I developed a new method to clone the suppressors combining whole genome sequencing/SNP mapping and RNAi screening. I also helped developed reverse genetic techniques using CRISPR/Cas9 and used these to verify the identity of one suppressor. My thesis demonstrates how new genetic tools can be used to facilitate the identification and study of suppressors in C. elegans. The methods I describe here could be applied to other biological questions and models.