Description:
This workshop is designed to focus on applications of CRISPR technology as a platform for genome editing and functional genomics. The program will provide a hands-on laboratory experience demonstrating targeted mutagenesis in cultured human cells using the Cas9:gRNA ribonucleoprotein (RNP) complex. In addition; the workshop will address topics in genome editing and CRISPR-Cas9 research, including basic and enhanced CRISPR methods, cellular repair mechanisms, regulation of gene expression, bioinformatics, applications to various organisms.
Topics that will be covered:
The intensive workshop is specifically designed to address
• gRNA synthesis using PCR and in vitro transcription (IVT).
• Purify gRNAs and assemble with Cas9 to form an RNP
• Nucleofect Cas9:gRNA RNP into cultured human cells using a lipid-based transfection reagent to generate a gene knockout.
• Determination of general editing efficiency with a PCR-based DNA mismatch method (genomic cleavage detection assay) and detection of the SNP change through analysis of fluorescent protein expression.
Duration: 5 working days (3 hours/day).
Course structure: The course will constitute theoretical classes, demonstration and hand training.
Workshop Agenda:
1st day: Overview of basic principles of CRISPR-Cas9 editing technology.
2nd day: Basic consideration for guide RNA sequence design.
3rd day: Genome editing of E.coli using CRISPR-Cas9 technique, highlight on the main difference between knock-in and knock out principle
4th day: Transfection of the Nucleofect Cas9:gRNA RNP into cultured human cells using lipoefectamine.
5th day: Determination of editing efficiency using PCR-based DNA mismatch method (genomic cleavage detection assay) and protein characterization.
Who Should Attend:
The workshops were designed for both beginners and those who are already working with the techniques but wishing to learn appropriate technical skills that can be applied in their practical field. Moreover; to learn and trained on new era of research technology.
Who will attend?
• Master/Ph.D. Students, Scientists
• Researchers
• Research Assistants, Technical Officers
• Academic Staff
• Undergraduate students of medical and para-medical specialities.
Supplements:
At the end of the workshop; attendees will receive the following documents:
• Official training Certificate (Signed & stamped).
• Full workshop review (PDF).
• Technical data troubleshooting memo (PDF).
• In addition, the participants will have a chance to win a prize on participation of a ‘quiz’ based on the knowledge gained during the workshop