940342 Molecular genetics of filamentous fungi - basic course

Type
Lecture and exercise
Semester hours
2
Lecturer (assistant)
Strauss, Joseph , Svoboda, Thomas
Organisation
Offered in
Wintersemester 2023/24
Languages of instruction
Englisch
Content

This newly designed laboratory course „Molecular genetics and genomics of hyphal fungi – basic course“ features education on [1] the phenotypic and molecular analysis of genetic crosses and [2] genome editing experiments using CRISPR-Cas9 for strain improvement and synthetic biology.

Detailed description

[1.]: Molecular analysis of markers segregating in sexual crosses. Different strains containing selectable and non-selectable markers will be crossed and the progeny will be analyzed phenotypically for the selectable markers and genotypically using PCR-amplification methods to follow segregation of non-selectable markers. Both, free segregation as well as recombination events will be analyzed. Theoretical background on mutation, selection and forwards genetic screens is presented and discussed.



[2] Construction of fungal strains by transformation that receive the genes encoding the CRISPR-Cas9 genome editing molecular machinery. The model organism Aspergillus nidulans will be transformed with a vector that contains both the Cas9 enzyme and genes coding for the single-guide-RNA (sgRNA). In a bioinformatic part the design of the sgRNAs for one or more loci that should be edited by cas9 will be trained. As a result of the successful transformation the targeted locus will be altered in order to change the capacity of this strain to produce certain bioactive metabolites (e.g. penicillin). The alteration of the metabolite levels will be tested in bioactivity assays in the follow-up course for advanced fungal genomic studies.
Previous knowledge expected

Solid knowledge of formal and molecular genetics and basic education in microbiological and molecular biological techniques
Objective (expected results of study and acquired competences)

Students gain insights into the theoretical background and experimental methods of molecular and formal genetic analyses and fungal strain improvements. A deeper insight into meiotic processes, recombination and offspring analysis will be gained. Environmental pollution with hormones and endocrine disruptors will be deeply discussed and a fungal bioreporter will be employed practically in the lab to detect these substances. Novel CRISPR-Cas9 genome editing technology will be discussed at the theoretical level and bioinformatic and practical exercises will reveal the relevance of this new technique for biotechnological and agro-genetic strain improvement.
You can find more details like the schedule or information about exams on the course-page in BOKUonline.