Why Humanize Mouse Genes
Species Specific Therapeutics
Many therapeutic antibodies and biologics are designed specifically for human targets:
- Therapeutic antibodies may not cross react with mouse orthologs
- Human specific epitopes require human sequence for binding
- Biologics may have species specific receptor interactions
- Humanized models enable efficacy testing of clinical candidates
Translational Relevance
Humanized models improve translation to clinical outcomes:
- Study human specific protein functions
- Model human specific disease mechanisms
- Evaluate pharmacokinetics with human target engagement
- Develop and validate human specific biomarkers
Regulatory Considerations
Humanized models support regulatory submissions:
- Demonstrate on target efficacy with human target
- Evaluate safety in context of human target biology
- Support mechanism of action characterization
- Enable pharmacodynamic biomarker development
Humanization Strategies
Complete Gene Replacement
The entire mouse gene is replaced with the human ortholog:
- Human coding sequence under mouse regulatory control
- Preserves mouse expression pattern and level
- Enables study of all human specific protein features
- Most comprehensive humanization approach
Complete replacement is appropriate when the entire protein sequence differs functionally between species or when studying human specific splice variants.
Extracellular Domain Humanization
Human extracellular domain replaces mouse sequence while retaining mouse intracellular domain:
- Human epitopes for antibody binding
- Mouse signaling domains for normal downstream function
- Commonly used for immune checkpoint humanization
- Maintains compatibility with mouse immune system
This approach is ideal for therapeutic antibody testing where the antibody targets extracellular epitopes.
Specific Domain or Epitope Humanization
Only the specific region recognized by the therapeutic is humanized:
- Minimal sequence change
- Preserves most mouse protein function
- Targeted to specific antibody epitope
- May require epitope mapping data
Humanized Regulatory Elements
Human promoter or enhancer sequences replace mouse regulatory regions:
- Study human specific gene regulation
- Model human expression patterns
- Investigate human specific enhancer variants
TruHumanization™ Technology
ingenious targeting laboratory's proprietary TruHumanization™ technology optimizes humanization strategies for maximum therapeutic relevance and physiological compatibility with complete gene replacement.
Quality Assurance: All TruHumanization™ projects include pre-germline analysis to verify correct human sequence integration before mouse generation.
Applications by Therapeutic Area
Immuno Oncology
Humanized immune checkpoint models are essential for cancer immunotherapy development:
Dual humanized models (e.g., PD1 + CTLA4) enable combination immunotherapy evaluation.
Learn moreMetabolic Disease
Humanized metabolic targets support therapeutic development:
- Humanized receptors for peptide therapeutic testing (GLP1R, GCGR)
- Humanized enzymes for small molecule drug development
- Humanized lipoproteins for cardiovascular studies
- Humanized transporters for pharmacokinetic evaluation
Neuroscience
Humanized CNS targets enable neurotherapeutic development:
- Humanized amyloid precursor protein (APP) for Alzheimer studies
- Humanized tau (MAPT) for tauopathy research
- Humanized receptors for antibody or biologic testing
- Humanized ion channels for species specific pharmacology
Infectious Disease
Humanized receptors enable infection and therapeutic studies:
- Humanized viral entry receptors
- Humanized immune components for vaccine studies
- Humanized targets for antiviral therapeutic testing
Strain Background
Choose strain background based on experimental requirements:
C57BL/6
Most common, compatible with syngeneic tumor models (MC38, B16)
BALB/c
Alternative background, compatible with CT26, 4T1 tumors
Immunodeficient backgrounds
For human immune cell engraftment studies
Combining Multiple Humanizations
Complex models may require multiple humanized genes:
- Dual checkpoint humanization (PD1 + CTLA4, PD1 + LAG3)
- Receptor ligand pairs (CD47 + SIRPα)
- Pathway components (multiple targets in same pathway)
Breeding strategies or sequential targeting can combine multiple humanized alleles.
Humanized Checkpoint Models
| Target | Humanization Approach | Applications |
|---|---|---|
| PD1 (PDCD1) | Extracellular domain | Anti PD1 antibody efficacy |
| PDL1 (CD274) | Extracellular domain | Anti PDL1 antibody efficacy |
| CTLA4 | Extracellular domain | Anti CTLA4 antibody efficacy |
| LAG3 | Extracellular domain | LAG3 inhibitor evaluation |
| TIM3 | Extracellular domain | TIM3 pathway studies |
| TIGIT | Extracellular domain | TIGIT inhibitor development |
| CD47 | Extracellular domain | Anti CD47 antibody efficacy |
| SIRPα | Extracellular domain | CD47 SIRPα axis studies |
What Researchers Say
“We are very happy with this mouse line and we're not done yet. There's still plenty to do. The versatility of the F.A.S.T.™ approach is really unparalleled. There are so many possibilities to use the cassette that one paper does not showcase it all. Also, the addition of N-terminal GFP has helped quite a bit to understand gene expression patterns.”
— Frank Bosmans, PhD
Ghent University, Belgium
Start Your Humanization Project
Our scientific consultants are ready to discuss your humanization requirements and recommend the optimal strategy for your therapeutic program. Initial consultation is provided at no charge and includes humanization approach recommendations, allele design options, and timeline estimates.
Key Terms
Understanding the terminology used in humanized mouse model generation helps you communicate effectively with our scientific team and interpret project documentation.
Humanized Mouse
A mouse model where a mouse gene or genomic region has been replaced with the corresponding human sequence, enabling study of human-specific biology.
Gene Replacement
A targeting strategy that substitutes mouse genomic sequences with human equivalents to model human-specific biology or test human-targeted therapies.
BAC (Bacterial Artificial Chromosome)
A DNA construct used for cloning large DNA fragments (100-300 kb) in bacteria, used for large-scale humanization projects.
Chimera
An organism composed of cells derived from two or more different sources, used to establish germline transmission of targeted alleles.
Humanization Resources
Download our free guides to help plan your humanized mouse model project.
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Frequently Asked Questions
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