TRAC 12:Transgenic Technology: Methods and Application
This lecture/laboratory demonstration course is intended for those who wish to develop a strong background in transgenic techniques and applications. The two dominant approaches to transgenic technology will be emphasized, namely classic transgenic production by pronuclear microinjection and knockout transgenic production(gene targeting) by blastocyst injection with embryonic stem (ES) cells. The comprehensive coverage will address supporting technology (e.g. cryopreservation, rederivation, Intracytoplasmic Sperm Injection(ICSI), In Vitro Fertilization (IVF) , animal husbandry and surgery) as well as the molecular biology/recombinant DNA aspects of transgenesis.
Topics: Transgenic Technology overview (Embryonic Development, Optimizing Constructs for in vivo Expression, Preparation of Females for Embryo Collection, Pronuclear Microinjection, Generation of Pseudopregnant Females, Implantantation in Foster Mothers, Identification of Transgenic Progeny, Optimization of Breeding Population from Founders); Lentiviral and BAC transgenisis; Gene Targeted Transgenisis (Isolation and Maintenance of Totipotent embryonic Stem Cells for the Generation of Chimeric Mice; Homologous Recombination in ES Cells; Construct Design, Transfection, Selection; Generation of Gene Knockout Mice using Targeted ES Cells; Blastocyst Injection; Optimizing Germ-line Chimeras); Alternative Methods for Generating Targeted Mice: 8_cell Embryo Injection and ES Cell Aggregation; transgenic phenotype analysis; Transgenic Animal Model Supporting Techniques (Cryopreservation and rederivation; ICSI and IVF; Transgenic Model Future Prospects); Transgenic Model FuFuture Prospects; Sampling Project Question and Answer; Live and Video Demonstrations and hands on work (e.g. microinjection; Superovulation; Embryo Harvest and Transfer).