Genetic Mice

4 Pages 1039 Words

Transgenic Outline
Introduction
Experimental Timeline
Plan the Experiment
Clone the Transgene
Transgene Screening
Transgene Expression
DNA Microinjection
Identify Transgenic Founders
Southern Analysis
Transgenic Analysis
Pertinent Publications
Review Articles and Books





1. Introduction. This is a brief outline of the steps necessary to obtain a transgenic mice. Simply put, the investigator constructs a transgene with a promoter and a structural gene (for example a reporter gene such as lacZ or a transcription factor). DNA is prepared and microinjected into fertilized mouse eggs. Potentially transgenic mice are born. Transgenic founders are identified and bred to produce offspring for analysis. Core personnel are available for consultation on all aspects of transgenic research. Contact Thom Saunders with any questions or Meet the Staff.

2. Plan the experiment. What is the purpose of your experiment? Do you want to analyze tissue specific regulatory sequences? Do you want to overexpress a protein in a specific cell lineage? You will need to obtain or clone the desired promoter and structural gene. Certain genes will be deleterious or incompatible with proper growth and development of the embryo, special arrangements should be made if you expect embryonic lethality from transgene expression. The expression of a transgene requires that the appropriate transcriptional control element be included in the DNA construct. Preliminary studies in cell cultures are recommended to verify the integrity of the construct and the function of the promoter. However, it is not always possible to predict in advance whether the transgene will have the capability of being expressed in vivo. The Core has a nuclear localized lacZ reporter vector (pnlacf) for investigators who wish to characterize regulatory elements in transgenic mic!
e. A completed materials transfer agreement is required before this plasmid can be...