Course

Unit 1

Unit 1:

Introduction to Genome Editing and the CRISPR-Cas System, History of genome editing technologies: ZFNs, TALENs, and CRISPR, Discovery and evolution of CRISPR-Cas systems, Structure and function of CRISPR-Cas9, Cas12, and Cas13, Types of CRISPR systems and their biological roles in prokaryotes, Comparison of CRISPR with other genome editing tools.

Unit 2

Unit 2:

CRISPR-Cas Mechanism and Design Strategies, Molecular mechanism of CRISPR-Cas9-mediated DNA cleavage, Guide RNA (gRNA) design and synthesis, PAM sequences and target site selection, CRISPR efficiency and off-target prediction, CRISPR toolkits: plasmid-based, ribonucleoprotein (RNP), and viral delivery.

Unit 3

Unit 3:

Delivery Systems and Experimental Implementation, In vitro and in vivo delivery methods: lipofection, electroporation, viral vectors, Cell types and model organisms used in CRISPR experiments, Genome editing workflows: knockout, knock-in, base editing, and prime editing, Validation techniques: PCR, sequencing, and functional assays, Troubleshooting and optimization strategies.

Unit 4

Unit 4:

Applications in Medicine and Biotechnology, Therapeutic applications: monogenic disorders, cancer, infectious diseases, CRISPR in diagnostics (e.g., SHERLOCK, DETECTR), Agricultural and environmental applications, Drug discovery and functional genomics using CRISPR screens, Case studies of clinical trials and preclinical research.

Unit 5

Unit 5:

Ethical, Legal, and Social Implications (ELSI), Germline vs somatic editing: ethical considerations, Regulatory landscape: global perspectives (FDA, WHO, etc.), Biosecurity and misuse concerns, Intellectual property and patent issues, Public perception and engagement in gene editing.

Unit 6

Unit 6:

Hands-On Laboratory Training and Project Design, Designing CRISPR experiments: target selection and gRNA construction, Performing CRISPR edits in model systems (e.g., bacteria, mammalian cells), Analyzing editing outcomes: genotyping, sequencing, and phenotyping, Group-based mini-projects: planning, execution, and presentation, Interpreting results and proposing future directions.

(45 classes)

Preamble:

This specialized course focuses on the CRISPR-Cas-based genome editing system, detailing its mechanisms, design strategies, and diverse applications in medicine. Discussions will also address the ethical, legal, and social implications of gene editing technologies. Laboratory sessions provide hands-on experience with CRISPR tools, enabling students to design and implement genome editing experiments effectively.

Course Outcomes:

CO1: Describe genome editing technologies with a focus on CRISPR-Cas systems.

CO2: Design and assess guide RNAs and CRISPR components for gene editing.

CO3: Apply CRISPR delivery methods and editing techniques in experimental setups.

CO4: Analyze editing outcomes using molecular and functional assays.

CO5: Discuss CRISPR applications in medicine, diagnostics, and biotechnology.

CO6: Evaluate ethical, legal, and social aspects of CRISPR and conduct project-based experiments.

Program outcome

PO1: Bioscience Knowledge

PO2: Problem Analysis

PO3: Design/Development of Solutions

PO4: Conduct Investigations of complex problems

PO5: Modern tools usage

PO6: Bioscientist and Society

PO7: Environment and Sustainability

PO8: Ethics

PO9: Individual & Team work

PO10: Communication

PO11: Project management & Finance

PO12: Lifelong learning

0 – No affinity; 1 – low affinity; 2 – Medium affinity; 3 – High affinity

Program-specific outcome

PSO 1 – Emerging technologies in cell therapy

PSO 2 – Biomolecules in Medicine

PSO 3 – Molecular dysregulation in diseases

PSO 4 – Molecular technology in diagnosis and therapy

PSO 5 – Applying lab discoveries to clinical practice

PSO 6 – Advanced techniques in cell culture, gene editing, and bioprocessing

PSO 7 – Statistical methods to interpret and validate diagnostic results

PSO 8 – Integrate cell therapy into personalized medicine

PSO 9 – GMP and regulatory practices in cell therapy production

PSO 10 – Bioinformatics and biological data use

0 – No affinity; 1 – low affinity; 2 – Medium affinity; 3 – High affinity

Textbook

Santisteban, M. S. (2023). Introduction to CRISPR-Cas9 Techniques. Springer.

Gasiunas, G., Barrangou, R., Horvath, P., & Siksnys, V. (Eds.). (2021). Introduction to CRISPR-Cas9 Techniques. Springer.

Reference

Synthego. (n.d.). CRISPR 101: Your Guide to Understanding CRISPR. Synthego Corporation.