📚 Syllabus & Course Curriculum
Biotechnology & Genetic Engineering
Module-by-module breakdown of A Hands-On Guide Using CRISPRdirect and CHOPCHOP, from foundations to a certified capstone project.
Setup
• Knockout, knock-in and tagging goals dictate entirely different guide placement
• Retrieving the correct transcript and genome build — a mismatch invalidates the design
• Exon choice for a true null: early constitutive exons, avoiding alternative starts
CRISPRdirect
• Running a target sequence and reading the returned candidate list
• Interpreting the 20mer, 12mer and 8mer uniqueness columns
• Species and genome selection as the most common source of a wrong answer
CHOPCHOP
• On-target efficiency scoring and the models behind the numbers
• Off-target counts by mismatch number and the weighting between them
• Nuclease choice, PAM options and restriction-site output for screening
Judgement
• Trading predicted efficiency against off-target risk for your application
• GC content, poly-T terminators and secondary structure in the sgRNA
• Why two or three independent guides should always be tested, not one
Validation
• Primer design for the amplicon and its distance from the cut site
• T7E1 and TIDE or ICE for editing efficiency estimation
• Sanger and amplicon sequencing to characterise the actual allele obtained
Earn government-registered certification in A Hands-On Guide Using CRISPRdirect and CHOPCHOP
e-Certificate and e-Marksheet issued on successful completion.
