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"I’ve been working with iTL over the past 5 years in the production of 3 different genetically altered mice. Not only did iTL help in the design of the mice, […]” - Raghu Mirmira, MD, PhD University of Chicago.
When it comes to understanding the significance of floxed mice, Cre lox recombination and similar methods for gene targeting are essential. This technique was created as a type of site-specific recombination process in order to delete or activate loxP-flanked DNA sequences. It is mainly used to define the roles of various genes and determine how they behave and how they can be influenced in relation to various genetic disorders and genetically influenced health complications.
Floxed mice, Cre recombination experts believe, have the best chance at offering relevant and useful data that can be adapted to human research later on. This is why they are the main types of animals used in experiments involving the loxP flanking of chromosomal DNA sequences. At first, site-specific recombination was used by Dr. Brian Sauer in his experiments to show it’s possible to activate specific gene expression in mammalian cell lines. Later, Dr. Jamey Marth demonstrated that Cre-lox could also be used on specific cell types, to define endogenous gene functions and determine the roles that genetic lesions played in various diseases.
In 1994, Drs. Marth and Rajewsky discovered that the Cre-lox system could be used for conditional gene targeting. Together with the work of Joe Z. Tsien, who came up with ways of applying Cre-lox to cells and region-specific genes for manipulation purposes, they pioneered the widespread use of conditional mutagenesis throughout the growing world of genetic research. Although Cre-lox recombination is a method developed less than 30 years ago and may seem relatively new, that is seen as quite a long period in the world of genetic research. Despite this, Cre-lox recombination is still regarded as a very precise method used for specific gene targeting.
When it comes to discussing floxed mice, Cre lox recombination techniques are often mentioned in the same sentence. This is because floxed mice, or mice with cells that bear a targeted loxP-flanked (also known as floxed) polymerase gene, are essential to ensure that the technique actually functions as intended. In Cre-lox recombination, the floxed gene is used to pinpoint the precise gene targeted for the recombination process. Once flanked at the short target DNA sequences known as the lox sequences, the piece of genetic material that is meant to be deleted, inverted, translocated or inserted can then interact with the Cre recombinase enzyme for recombination to occur.
With the help of floxed mice, Cre lox recombination processes can be applied to splice snippets of DNA in ES cells in order to generate mice with specific genetic enhancements or detriments. Genes can not only be added, deleted or inverted, but they can also be partially inactivated for the purpose of studying how a faulty gene might impact the health and development of an animal model with DNA similar to that of a human. Modern advancements have taken the process to the next level, continuing to use floxed mice, Cre recombination techniques and new technologies that obtain accurate data more quickly than before.
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