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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.
The gene knockout process is one of the most important genetic techniques used for altering the genetic makeup of mammalian creatures. Typically used on mice, it involves a number of complex processes and methods that can be used for facilitating the inactivation of a gene. Whether we’re talking about conditional or regular gene knockouts, or about advanced methods such as CRISPR/Cas9 and TALENs, the method can be considered one of the most effective when it comes to the study of human disease.
Some techniques for manipulating genes are used to modify or add genetic material to the genome of a certain organism. Others are used to remove or deactivate certain genes. The gene knockout is a method that falls under the latter category. It typically involves the use of an animal model such as a laboratory mouse (also known as a knockout mouse) and a tool such as zinc fingers, homologous recombination, or the CRISPR/Cas9 technique for obtaining knockouts. There are different types of knockouts depending on the type of gene that’s being targeted, the conditions involved with the method, or the number of knockout genes. Double, triple and even quadruple knockouts are all possible. One also has to distinguish between homozygous and heterozygous knockouts.
Knocking out genes in the mammalian genome has been done for quite a few years, and the methods involved are being perfected to an increasing extent almost on a yearly basis. Today there are countless universities and scientific institutions, as well as amateurs, who attempt to create new KOs using established gene knockout methods. Recently, the International Knockout Mouse Consortium has expressed the goal to knock out every single gene in the mouse genome, and they have already started. A recent paper published by Karen Svenson, Robert Braun and Stephen Murray has already documented the detailed analysis of the first 3,328 genes.
A gene knockout is essentially a tool that’s used in order to help researchers understand the role of a specific gene or a certain portion of DNA. By “knocking out” or deactivating a certain gene, the method allows scientists to study the process by which disease sets in as a result of the deactivation or impairment of certain genes. When the gene is knocked out, the organism starts exhibiting symptoms resulting from gene impairment. Researchers then have the opportunity to use various methods designed to solve the problem. Knockouts are, therefore, frequently used as disease models that help geneticists better understand the origins of certain diseases.
Our understanding of human health and disease has been advanced exponentially since knockouts began appearing on the scene. The first gene knockout was developed more than 30 years ago, and the process is still being perfected today. Since then, countless genetic models of human disease have been developed through humanized mice and research conducted directly on embryonic stem cells. Whether naturally occurring or a result of laboratory tests, humanized knockout mice have become an important tool for studying genetic diseases like Krabbe disease and muscular dystrophy. They’re also used in studying cancer, diabetes, Alzheimer’s and many other well-known disorders that can be linked to genetics, but only develop under certain circumstances.
With the aid of knockout mice, genetic researchers have not only made significant breakthroughs in understanding the onset of certain diseases, but in treating them as well. Humanized mice help to avoid the need to use human subjects in studies that would normally be dangerous or unethical. Moreover, with the help of humanized mice, researchers are better able to understand the function of genes as they relate to each other. As a result, the use of a single or multiple gene knockout can often lead to unparalleled discoveries that would normally be difficult to attain.
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