Transgenic animal technology : a laboratory handbook / edited by Carl A. Pinkert.

Section One Overview. Introduction to transgenic animals. Section Two Transgenic animal production focusing on the mouse model. Transgenic animal production using DNA microinjection. Factors affecting transgenic animal production. Gene targeting in embryonic stem cells I: History and methodology. Gene targeting in embryonic stem cells II. Conditional technologies. Retrovirus-mediated gene transfer. Nuclear transfer technologies. Section Three Production of transgenic laboratory and domestic animal species. Production of transgenic rats. Production of transgenic rabbits. Production of transgenic fish. Production of transgenic poultry. Production of transgenic swine by DNA Microinjection. Production of transgenic ruminants by DNA Microinjection. Production of transgenic non-human primates. Retrovirus-mediated gene transfer into oocytes. Section Four: Molecular biology, analyses and enabling technologies Vector design for transgene expression. Analysis of transgene integration. PCR optimization for detection of transgenic integration. Evaluation of transgene expression. Control of gene expression (from standard gain/loss of function to anti-sense to PNAs). Enabling technologies: Cryopreservation of gametes. Enabling technologies: Embryo culture considerations. Databases, Internet Resources and Genetic Nomenclature.

Front Cover; Transgenic Animal Technology; Copyright Page; Contents; List of Contributors; Preface; One: Overview; 1 Introduction to Transgenic Animal Technology; I. Introduction; II. Historical Background; III. Applications and Overview of Text; References; Two: Transgenic Animal Production Focusing on the Mouse Model; 2 DNA Microinjection, Embryo Handling, and Germplasm Preservation; I. Introduction; II. General Methods; A. DNA Preparation and Purification; 1. DNA Construct/Fragment Structure; 2. Factors that Influence Transgenic Mouse Production; 3. Preparation of DNA for Microinjection.

A. Construction and Isolation of DNAb. DNA Purification; c. Purification and Quantification of DNA; B. Superovulation, Egg Culture, and Harvest; 1. Mouse Strains for Microinjection and Transgenic Models; a. Strains; b. Genetic Variability and Transgene Expression; 2. Superovulation of Mice; a. Pregnant Mare Serum Gonadotropin and Human Chorionic Gonadotropin; b. Follicle Stimulating Hormone and Luteinizing Hormone Delivered with Osmotic Pumps; 3. Production of Eggs from Superovulated Females; a. Colony; b. Influence of Litter Number; 4. In Vitro Culture of Superovulated Eggs; a. Equipment.

B. Atmosphere for Culturing Eggsc. Glass Pipettes for Manipulating Eggs; d. Media; e. Water Sources; f. Culture Dishes; 5. Harvesting Superovulated Eggs; a. Mating; b. Oviduct Dissection; c. Release of Eggs from the Oviduct; d. Egg Development and Microinjection Timing; e. Efficiency of Egg Culture; C. Microinjection Needles and Slides; 1. Types of Glass; 2. Preparation of Glass; 3. Pipette Pullers; 4. Injection Pipette Shapes; 5. Beveling Hollow Capillary Microinjection Needles; 6. Microforge Shaping of Pipettes; a. Manufacturers; b. Heating Filament; c. Holding Pipette; d. Injection Pipette.

7. Microinjection Slide for Eggsa. Slide Designs; b. Microinjection Media; c. Arrangement of DNA and Media Drops; D. Microinjection Equipment; 1. Microscopes; 2. Micromanipulators; a. Micropipette Holders; b. Micromanipulator Adjustments; c. Antivibration Tables; 3. Microinjection Systems; a. General; b. Oil-Driven Injection Assembly; 4. Oil-Driven Holding Pipettes; 5. Electronic Delivery Systems; E. Microinjection Procedure; 1. Preparation of Injection and Holding Pipettes (Micrometer Syringe System); a. Holding Pipette; b. Injection Pipette; c. Aligning the Injection Needle.

D. Loading DNA into the Injection Needle2. Preparing Eggs for Microinjection; 3. Steps for Efficient Manipulation of Eggs During Microinjection; a. General; b. Micromanipulators; c. Microinjection Needle; d. Controls and Egg Manipulation; 4. Visualizing and Injecting the Egg Pronuclei; a. Orientation; b. Egg Membrane Penetration with the Injection Needle; c. Filling the Pronuclei with DNA Solution; 5. Criteria for Microinjection to Yield Transgenic Mice; a. Treatment After Microinjection; b. Egg Lysis; c. Record Keeping; d. In Vitro Culture of Microinjected Eggs; F. Egg Transfer.

Transgenic animal technologies and the ability to introduce functional genes into animals have revolutionized our ability to address complex biomedical and biological questions. This well-illustrated handbook covers the technical aspects of gene transfer - from molecular methods to whole animal considerations - for important laboratory and domestic animal species. It describes methodologies as employed by leading laboratories and is a key resource for researchers, as well as a tool for training technicians and students. This second edition incorporates updates on a variety of genetic engine.

Description based on print version record.