The use and study of transgenic organisms raises legal issues, particularly the potential for patent infringement if a necessary license is not obtained. Scientists, regardless of whether they practice in an academic or corporate setting, should be aware of patents relating to the resear ...
Transgenic mouse models open new avenues of research to understand gene function, to mark cells, and to model human genetic diseases. The use of large DNA transgenes provides more information to cells and tissues in the mouse so that gene expression occurs at physiological levels in the appropr ...
The use of genomic-type DNA constructs ensures optimal transgene expression, once inserted into the host genome, because their large size includes most if not all the regulatory elements that are needed for correct gene expression. Large heterologous DNA molecules can be easily manipul ...
The rat is an important system for modeling of human disease. The use of transgenesis is relatively uncommon in rats. In this chapter, we focus on describing efficient techniques for the generation of transgenic rats by microinjection of plasmid DNA into pronuclei and the injection of human imm ...
The introduction of a transgene into a fertilized oocyte by pronuclear microinjection of a solution containing the construct of choice is probably the most straightforward method to generate a genetically modified organism. This technique has been adapted to a number of vertebrate and ...
The efficiency with which transgenic mice can be produced via pronuclear injection of DNA constructs is subject to a large number of variables ranging from human to mechanical to biological. Transgenic core facilities, which are often run like small businesses that must attract and satisfy ...
The use of genetically engineered mice to understand gene function is widespread. Changes to the mouse genome can be introduced with gene targeting vectors or with transgenes. Targeting vectors are usually used to ablate gene expression while transgenes are designed to express proteins ...
In theory, designing a DNA construct to be used for transgene purposes, for standard pronuclear microinjection, would seem a rather easy task. The combination of a given promoter and some regulatory elements of choice, driving the expression of the construct to the desired tissue, with a suitab ...
The design of any new mouse genetic modification today should start with careful scrutiny of the resources that are already available, through the internet, for information relating to your gene of interest. International mouse consortia are constantly providing new genetically mo ...
In their 1959 publication The Principles of Humane Experimental Technique, Russell and Burch defined three criteria to be used to alleviate the sources and incidences of inhumanity when performing animal experimentation. These include reduction, replacement, and refinement. To ...
The increased popularity of genetically modified animals in collaborative studies has stimulated the widespread interchange of mice among institutions with inconsistent health standards. While the presence of certain organisms may be tolerated at one institution, the impact ...
Following the advent of transgenic technology and the subsequent establishment of international consortia for broad-based genomic analysis of the mouse, the need for sharing of resources in the exchange of mouse models and material has reached an all time high. Live animals, embryos, and g ...
Cryopreservation is the method of choice not only for archiving mouse mutant models at the end of an experiment, but also to create a security back up during their development. Three key factors need to be considered in cryopreservation: why, when, and how. In this chapter, we intend to discuss these fac ...
Technical advances in the generation of genetically modified (GM) mice and the efforts of large-scale consortia have provided a wealth of resources to the biomedical research community. It has never been easier to obtain a specifically modified allele. However, the establishment and pr ...
The techniques of pronuclear and blastocyst injection are now routine procedures for making transgenic and gene-targeted mice, respectively. For many institutions, Core Facilities are the standard venue for producing such genetically modified animals for use in both basic and bio ...
The ability to directly reprogram mammalian adult somatic cells to an undifferentiated pluripotent stage similar to that of embryonic stem cells by introduction of a reduced number of transcription factors has opened new venues in many fields of Biology and Medicine. These reprogramm ...
Embryonic stem (ES) cells have had a tremendous impact on the field of genetics and are widely used as a means for precise genetic modification of the mouse genome. This chapter will give a background to the use of these versatile cells and provide practical protocols for their derivation and culture. ...
The marvel of embryonic stem (ES) cells is that after in vitro culturing and genetic modification, they still have the ability to contribute to the developing embryo, when combined with pre-implantation embryos, to produce chimeras and even completely ES cell-derived animals. In this chap ...
The generation of mouse chimeras by combining host embryos with genetically modified embryonic stem (ES) cells is a necessary step towards establishment of genetically modified mouse strains. This protocol describes the procedures necessary for the production of mouse chimeras by ...
The choice of a specific parental ES cell line used to create a genetically modified mouse has critical impact on the overall success of the project – affecting costs, complexity of effort, and time to efficient project completion. Despite the importance of making a thoughtful choice, many people ...
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