A point mutation is a genetic mutation in the form of insertion/deletion (indels) or substitution to a single nucleotide base in the genome sequence, resulting in reduced or changed downstream protein function. Therefore, point mutation mouse models are generated to study the roles of particular nucleotides and to model human genetic diseases.
Generally, point mutation models can be generated permanently, so-called conventional point mutation models, or in a time-controlled and tissue-specific manner, so-called conditional point mutation models. Creative Biolabs is capable of providing both conditional and conventional knock-in model creation services for our clients.
How to Create a Point Mutation?
Point mutations can be introduced by the approach of homologous recombination in ES cells. In this means, three steps are needed. First, the point mutation is inserted into one homologous arm of the gene-targeting vector. Second, mutant gene is introduced into the endogenous gene in the ES cells via homologous recombination. Third, the selection markers flanked by two loxP sites are removed by the Cre recombinase in the ES cells to create conventional models, or by breeding the mice produced from the targeted ES cells with another Cre-expressing mouse, to obtain time-controlled and tissue-specific models.
Conventional Point Mutation
A conventional point mutation results in the targeted allele encoding only the mutant sequence under the control of wide-type regulatory elements. It can be used as an alternative way of functional gene knock-out methods to avoid complex phenotypes. However, a conventional point mutation may exhibit lethal effects, in which case the inducible or conditional point mutation is needed.
Conditional Point Mutation
If you want more control over the expression of the mutant sequence, consider a conditional point mutation strategy. A conditional point mutation allows time-controlled and tissue-specific expression of the introduced mutated gene, which is achieved by using the LoxP/Cre system. For more information, please click the links below:
Besides the ESC/HR-based point mutation knock-in approach, point mutation mouse model generation can also be achieved using the latest CRISPR/Cas9 technology by co-injecting nuclease and oligo donors carrying the desired mutation and homology arms.
In the meanwhile, Creative Biolabs also offers other types of knock-in models:
Different strategies are needed for different experimental planning. Design the model for yourself and we'll do the ‘proof of concept' work, or consult Creative Biolabs' highly experienced technical team to determine the most appropriate strategy. At Creative Biolabs, you will get the best model creation service at the most favorable prices. Contact us or send us an inquiry for more information or a formal quote.
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