Transgenic Technology: Principles and Applications

Objective

This course is proposed for those who wish to develop a strong background in technologies and principals involved in the generation of genetically modified experimental organisms, from worms to animals, and explore their use in scientific research.

Syllabus

History and overview of transgenic technology- Molecular technologies used in transgenic technologyDirect single-cell embryo pronuclear injection, embryonic stem cells, and somatic cell nuclear transfer- Lentiviral and BAC transgenesis- Zinc finger nuclease technology- Genome editing approaches such as TAL effector nucleases and Cas9/CRISPR- Generation of the stable transgenic and mutant nematode worm Caenorhabditis elegans- Production of transgenic and mutant zebrafish and Xenopus- Production of transgenic mice (Embryonic development, Design, and optimizing Constructs for transgenic expression, Preparation of females for embryo collection- Pronuclear microinjection, Generation of Pseudo pregnant Females, Implantation in foster mothers, Identification of transgenic progeny)- Generation of knock-out mice ( Isolation and culture of embryonic stem (ES) cells, Gene targeting construct design, Transfection, Homologous recombination in ES Cells, Positive and negative selection; blastocyst injection, Breeding of germ-line chimeras)- Transgenic animal model supporting techniques (Cryopreservation and rederivation; ICSI and IVF)- Cre/lox and Flp/FRT system for inducible transgenics, general knockouts, conditional knockouts, and reporter strains in mice – Chemically inducible transgene expression systems- Use of transgenic technology in the modelling of human diseases, including cardiovascular diseases, diabetes, obesity, cancer, atherosclerosis, neurodegenerative diseases, muscle degeneration, and aging.