Tissue-specific point mutation models are defined as models in which a point mutation is introduced into the target gene of specific cells or tissues. Usually, the replacement of specific bases will result in mutated proteins and is used to study gene function. Creative Biolabs offers customized tissue-specific point mutation model creation services. Moreover, if it is combined with the induction system, both spatial and temporal control of the point mutation can be realized.
These conditional knock-in lines of mice result from a more sophisticated use of the existing gene targeting strategies that are used to create conditional knock-out models, such as the LoxP/Cre system. The difference is that the gene targeting vectors previously used to mediate the deletion of gene sequences are now crafted to mediate the one-for-one replacement of sequences so that a specific point mutation could be engineered into the genetic locus of interest.
Mechanisms of Tissue-Specific Point Mutation Models
In these models, conditional point mutation is achieved by using the site-specific recombinase system which is composed of two elements: the small repetitive sequence elements and the cognate recombinases. Specifically, on one hand, a line of mice with recombination sequence elements (LoxP or FRT) flanking the targeted endogenous gene is generated by using genetic manipulations. On the other hand, a line of mice with specific tissues expressing the cognate recombinase (Cre or FLP, respectively) is generated simultaneously. By crossing these two lines of mice, conditional point mutation mice are generated in which genetic recombination and point mutation occur only in the cells or tissues that express the necessary recombinase. By breeding with different Cre or FLP mouse, the mutation of the target gene in any tissue or cell of the mouse can be realized. Moreover, the expression level and profile of the mutated protein is accordant with that of the wild-type gene and is expressed during the whole development period.
Features of Tissue-Specific Point Mutations
Although this strategy is more complex than a global knock-in strategy (i.e. a conventional point mutation), complex phenotypes and even lethal effects could be avoided by creating lines of animals where the mutated gene is only expressed in specific cells or tissues. Meanwhile, this strategy allows very targeted and limited changes, thereby presenting useful tools for enhancing our understanding of gene function and its role in both health and disease.
Other knock-in strategies that you may be interested include:
Creative Biolabs has constructed systematic data and experience in generating tissue-specific and inducible point mutant rodent models. In order to meet your goals, a group of highly experienced scientists will be assigned to work closely with you to develop detailed study protocols.
Creative Biolabs is willing to apply our expertise in facilitating the brilliant studies of our clients. If you're interested in our services, contact us for more information or send us an inquiry for a formal quote.
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