Humanized mice are the perfect model system for studying human disease and for developing better therapeutics. A humanized mouse model can carry functioning human genes, cells, tissues, and/or organs. Boost the impact of your next project by replacing your gene of interest with the corresponding sequence from the human genome. Design your ideal humanized mice without limitation – from a single base change to replacement of a 200kb gene, including the potential for conditional alleles, cassette knockins, and more. Genetically humanized mouse models from ingenious will be the cornerstone of your research for years to come.
Change a single base or an entire gene to match the human sequence.
No design is too difficult – go beyond size limitations using our proprietary technology.
Create a single model for exploratory and translational research.
Full or Partial Gene Humanization
Replace Entire Coding and Noncoding Sequence Using ingenious’ TruHumanization™ Technology.
Traditional approaches to humanize the mouse manipulate only the coding regions of the genes of interest. Research has demonstrated that non-coding regions of the genome have implications in gene regulation and function. Therefore, it is becoming increasingly important to include non-coding regions of a human gene of interest when targeting the mouse gene ortholog.
There is no longer the limitation of leaving out human noncoding regions in your design.
Create a mouse model that better captures the human disease condition.
ingenious makes it easy to replace entire mouse genes with human.
Enhance your humanized mice by adding a reporter or making a conditional allele.
Let ingenious create the ultimate model for you by fully humanizing your gene of interest using TruHumanization™ technology. Up to 200kb can be replaced in a single step including coding and noncoding sequence. A humanized gene can also be made into a conditional allele for increased control over expression, and can incorporate known disease-causing mutations. Fully humanized mouse models lead to more faithful gene regulation and processing as well as the potential to perform functional and regulatory studies on multiple transcripts, providing you with a more flexible mouse model to study for years to come.
EXPRESS HUMAN CDNA FROM MOUSE LOCUS
- Expression of human coding sequence under the control of the mouse promoter.
- Can incorporate mutations, co-express reporter gene.
- Can include loxP design for tissue specific or temporal control.
- A cDNA knockin may be right for you if you need more than a simple point mutation.
Human cDNA can be knocked into any point of the mouse gene from the first ATG to the final exon. A cDNA at the 5’ end of the gene will be driven by the native promoter and will incorporate a poly-A sequence to block translation of the mouse gene. Add a fluorescent reporter to the knocked in sequence to track expression at the level of individual cells.
POINT MUTATION – CODING OR NONCODING
- Duplicate a mutation identified in human patients.
- Change a key amino acid to match human sequence.
- Conditional point mutations give you control over affected tissues.
ingenious has delivered hundreds of mouse models with point mutations. Contact us to discuss the right model for you, whether it be a point mutation in coding sequence, a promoter or a splice site. Your next model can more closely match the human sequence – wild-type or mutated.
ENHANCED CAPABILITIES AVAILABLE AT INGENIOUS
- Conditional activation of any humanized project by incorporating Cre-loxP technology into the design.
- Conditional activation of a human cDNA from the mouse Rosa26 locus, allowing more specific control of expression of the relevant human protein in the mouse model.
- Addition of F.A.S.T.™ cassette to enable inducible/reversible control of human gene.
- Addition of reporters or tags for co-expression with human protein.
- And more! No design is too difficult or out of reach when you work with ingenious’ experts.
Human Disease Model
The mouse has a long history as a model organism for the study of human disease and basic biology, based on the similar genetics and physiologies of the two organisms. In some cases a key difference between mice and humans can interfere with a particular experiment, for example a slight change in a protein domain or active site may interfere with antibody or small molecule binding. Differences at the tissue level can also be significant, preventing studies of systems such as the human immune system. For such cases mice can be “humanized” – altered at the genetic or cellular level to more closely match human biology. Several genetic humanization strategies are used to accomplish the goal of expressing human genetic sequences in addition to or in place of homologous mouse sequences, ranging from mutating a few key bases for a critical protein domain to replacing entire genes including noncoding regions. Gene replacement models are an exciting system enabling the study of splicing and other regulatory mechanisms in the context of a living animal. Tissue-level humanization is accomplished by xenografts, human tissues transplanted into mice lacking a fully-functioning immune system (to avoid having the mouse immune system reject the human cells). Genetically humanized models are more straightforward to use in experiments, if both strategies are an option for your research. Xenografted mice must be prepared individually, and the immune-compromised mice require special care. Genetically humanized mice pass on the changes to their offspring and the modifications can be made in healthier, wild-type genetic backgrounds. For more details contact ingenious today to discuss your next humanized mouse model.
Why create humanized mice?
Humanized mice are created to fill gaps in pre-clinical testing and have become an important tool for biomedical research. Animals are used as models of human biology due to ethical and logistic restrictions in using cells and tissues from human donors. In some cases the physiology of mice is not sufficiently close to human biology and the mice must be altered to create a better model system. Humanized mice are created in particular due to their ease of use, their sharing of genomic and physiological properties with humans, and their ability to be readily manipulated genetically.
RESOURCES FOR FURTHER READING
Church DM, Goodstadt L, Hillier LW, Zody MC, Goldstein S, She X, Bult CJ, Agarwala R, Cherry JL, DiCuccio M et al. 2009. Lineage-specific biology revealed by a finished genome assembly of the mouse. PLoS Biol 7: e1000112.
Devoy A, Bunton-Stasyshyn RK, Tybulewicz VL, Smith AJ, Fisher EM. 2011. Genomically humanized mice: technologies and promises. Nat Rev Genet 13: 14-20.
Macdonald LE, Karow M, Stevens S, Auerbach W, Poueymirou WT, Yasenchak J, Frendewey D, Valenzuela DM, Giallourakis CC, Alt FW et al. 2014. Precise and in situ genetic humanization of 6 Mb of mouse immunoglobulin genes. Proc Natl Acad Sci U S A 111: 5147-5152.
ENCODE (ENCyclopedia Of DNA Elements): The goal of this international database is to map functional elements within the human genome to the biological functions they regulate.
“We are 100% satisfied with iTL’s service in generating this mouse model for us. The model was completed quickly, with the utmost efficiency and each member of their team was extremely responsive and helpful. The full service guarantee, as well as the company’s excellent reputation were major decision factors for us.”
-Joseph A. Fraietta, PhD