Course

Unit 1

Unit 1:

Gene Editing with CRISPR-Cas9, Designing guide RNAs (gRNAs) for specific gene targets, Preparation and delivery of CRISPR-Cas9 components into mammalian cells, Screening and validation of gene edits using PCR and sequencing

Unit 2

Unit 2:

Transfection and Transduction Methods, Comparison of transfection techniques: chemical (lipofection), electroporation, and viral transduction, Optimization of transfection efficiency and expression analysis, Use of reporter genes and selection markers

Unit 3

Unit 3:

Functional Assays for Gene Expression, Quantitative PCR (qPCR) for gene expression analysis, Western blotting for protein expression and modification studies, Flow cytometry for cell surface markers and intracellular proteins

Unit 4

Unit 4:

Data Analysis and Interpretation, Statistical methods for analysing experimental data, Graphing and presentation of results, Trouble shooting common experimental issues

(45 classes)

Preamble

Cell and Gene Therapy Lab – II is an advanced laboratory course designed to provide students with hands-on experience in cell isolation, recombinant DNA technologies, expansion, cryopreservation, viability assays, and functional characterization methods used in therapeutic settings using flow cytometry.

Course Outcome:

CO1: Design and perform CRISPR-Cas9-based gene editing experiments, including guide RNA development and validation of gene modifications.

CO2: Compare and apply various transfection and transduction methods for gene delivery into mammalian cells.

CO3: Conduct gene and protein expression studies using qPCR, Western blotting, and reporter assays.

CO4: Utilize flow cytometry techniques for the analysis of cell surface and intracellular markers in engineered cells.

CO5: Apply statistical tools to analyse experimental results and generate meaningful conclusions from gene therapy experiments.

CO6: Troubleshoot common technical issues encountered in advanced gene and cell therapy workflows and optimize experimental design.

Program outcome

PO1: Bioscience Knowledge

PO2: Problem Analysis

PO3: Design/Development of Solutions

PO4: Conduct Investigations of complex problems

PO5: Modern tools usage

PO6: Bio scientist 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

• Scherman, D., Barratt, G., & Lautier, D. (2011). Advanced Textbook on Gene Transfer, Gene Therapy and Genetic Pharmacology: Principles, Delivery and Pharmacological and Biomedical Applications of Nucleotide-based Therapies. World Scientific Publishing.

Reference book

• Gen Script Biotech Corporation. (2021). Cell and Gene Therapy Handbook. Gen Script.