Detection methods for DNA changes caused by CRISPR/Cas9

Osrednja tema o kateri govorim v svoji diplomski nalogi so metode za detekcijo sprememb DNA povzročenih s CRISPR/Cas9. V uvodnem delu naloge opisujem razvoj odkritja naravnega CRISPR/Cas9 sistema skozi zgodovino. V nadaljnem delu sledi opis zgradbe CRISPR lokusa ter strukturne organizacijo in arhitekturo SpCas9, kjer sem se osredotočila na opis ključnih komponent za delovanje CRISPR/Cas9 sistema. V nadeljevanju sem predstavila način delovanja inženirskega CRISPR/Cas9 iz SpCas9 in njegovo uporabo kot molekularno orodje, s poudarkom na možnih spremembah v DNA in možne aplikacije. V drugi polovici diplomskega dela opisujem različne metode za detekcijo sprememb v DNA povzročenih s CRISPR/Cas9, možnosti njihove uporabe in prednosti ter pomankljivosti posameznih metod.The main topic that I am discussing in my thesis is about methods for detecting changes in DNA caused by CRISPR/Cas9. In the introductory part of the paper I describe the development of the natural CRISPR/Cas9 system through history. The following is a description of the structure of the CRISPR locus and the structural organization and architecture of the SpCas9 domain, where I focused on describing the key components for the operation of the CRISPR/Cas9 system. Then I introduced how the engineered CRISPR/Cas9 from SpCas9 works and its use as a molecular tool, with an emphasis on possible changes in DNA and possible applications. In the second part of the diploma thesis I described various methods for detecting changes in DNA caused by CRISPR/Cas9, the possibilities of their use, and the advantages and disadvantages of individual methods