📚 Syllabus & Course Curriculum
Biotechnology & Genetic Engineering
Module-by-module breakdown of Advanced Gene Editing of iPSCs Using CRISPR/Cas9, from foundations to a certified capstone project.
Cell Culture
• Feeder-free maintenance, passaging technique and colony morphology assessment
• Pluripotency verification and genomic stability monitoring by karyotype or SNP array
• Establishing a baseline before editing so later changes are attributable
Guide Design
• PAM constraints and on-target activity scoring
• Off-target prediction with Cas-OFFinder and CFD scoring, and the limits of prediction
• High-fidelity variants: eSpCas9 and SpCas9-HiFi
• When base editing or prime editing is the better tool than a double-strand break
Delivery
• RNP nucleofection versus plasmid and viral delivery in iPSCs
• NHEJ knockout versus HDR knock-in: choosing the repair outcome
• Donor template design: ssODN versus AAV, homology arm length, silent PAM mutation
• Cell-cycle timing and HDR enhancement, with realistic efficiency expectations
Screening
• Single-cell cloning and surviving the bottleneck: ROCK inhibition and plating density
• Editing efficiency deconvolution from Sanger traces using ICE or TIDE
• Zygosity determination and detection of large deletions that PCR can miss
• NGS amplicon validation of the final clone
Safety
• Empirical off-target discovery: GUIDE-seq, CIRCLE-seq and DISCOVER-seq
• Copy-number and karyotype reassessment after clonal expansion
• p53 pathway selection pressure and its implications for edited iPSC lines
Application
• Directed differentiation of edited lines to the relevant lineage
• Isogenic pairs: designing the comparison so the phenotype is attributable to the edit
• Functional assays and phenotype validation
• Reporting standards and line registration for reproducibility
Earn government-registered certification in Advanced Gene Editing of iPSCs Using CRISPR/Cas9
e-Certificate and e-Marksheet issued on successful completion.
