Next-Generation Humanized Mouse Model Construction Program

Launching the HUGO-GT™ Program:

Fostering Global Collaboration in Constructing Advanced Whole Genomic DNA Humanized Mouse Models, Catalyzing the Pace of Gene Therapy Drug Development.

Full Genomic Coverage of Pathogenic Gene

Efficient Drug Screening

Applicable to Gene Therapy Methods including ASO/CRISPR/siRNA, etc

HUGO-GT™ Whole Genomic DNA Humanized Model

The humanized mouse model offers the advantage of better recapitulating human physiological and pathological characteristics compared to traditional animal models, making it a preferred tool for studying human diseases and evaluating the efficacy and safety of potential therapeutics. This enhanced relevance to human biology allows for more accurate prediction of drug responses and disease mechanisms, potentially leading to improved translation from preclinical research to clinical applications.

However, most commonly used humanized models, such as transgenic (Tg) mice, coding sequence (CDS), and single-exon humanized mice, can only achieve partial insertion of the human gene into the mouse genome. These common, current-generation, humanized models exhibit notable limitations, including random insertion, complex genetic background and line establishment, as well as inadequate humanized regions.

To facilitate further advancement of disease mechanistic study and related drug development, full-length genomic DNA humanized mice which can faithfully recapitulate the expression pattern, regulation, and functional properties of the human gene in a mouse model, is urgently needed to replace the existing animal models and reshape the translational research framework for drug development. However, achieving entire genomic DNA sequence replacement poses technical challenges, as the large-scale introduction of exogenous sequences may affect the expression and regulation of endogenous genes, which brings a very daunting problem.

Driven by above demands, Cyagen has launched the HUGO-GT™ (Humanized Genomic Ortholog for Gene Therapy) program. We use our proprietary TurboKnockout-Pro technology to achieve in-situ replacement of the targeted mouse endogenous gene and successfully construct full-length genomic sequence humanized mouse models that encompass a broader range of intervention targets.

The HUGO-GT™ mice employ more efficient large-fragment vector fusion technology, which serves as a versatile template for targeted mutagenesis customization services, providing clinically relevant models that are closely aligned to real-world biological mechanisms.

Additionally, we offer CRO services in fields including ophthalmology, neuroscience, tumor immunology, and other disease areas, to empower research on genetic diseases and facilitate the development of gene therapy drugs.

Research Approach of the HUGO-GT™ Whole Genomic DNA Humanized Model

RDDC

TurboKnockout^®

Mature targeting technology, precise modification, reduces two generations of breeding time compared to traditional ES targeting technology

BAC fusion

By recombining, constructing larger BACs that facilitate the introduction of larger genes and regulatory sequences, thereby approaching the expression pattern of endogenous genes

RMCE

Achieves human-mouse gene replacement through the displacement function of recombinases

Target discovery

Target validation

Pharmacokinetics

Toxicity risk assessment

Collaboration Model of HUGO-GT™ Program

Internal Research

Model development
and technology commercialization

Collaborative Development

Licensing of whole genomic DNA humanized models

Novel drug development

Other flexible collaboration options

Join Us

Our HUGO-GT™ program encourages multiple collaborative models, including internal research, as well as open collaboration. If you are currently engaged in gene therapy research such as allele-specific oligonucleotide (ASO)/CRISPR/siRNA or have any other inquiries, please feel free to reach out and join our efforts in developing the whole genomic DNA humanized models that will best support translation of your preclinical studies.

Advantages of HUGO-GT™ Genomic DNA
Humanized Model

Applicable to a Variety of Preclinical Drug Experiments

In the pipeline of gene therapy for diseases such as Duchenne muscular dystrophy (DMD) and Retinitis pigmentosa (RP), there have been multiple successful cases of preclinical drug development using humanized disease models. The Cyagen HUGO-GT™ model not only serves to replace existing humanized models to meet the experimental research needs of most pharmaceutical companies, especially for those gene therapy drugs with high requirements for gene sequence integrity, such as ASO, CRISPR, and siRNA.

Enhanced Drug Screening Efficiency

As verified by our internal research team, the humanized region of the HUGO-GT™ model is consistent with the pathogenic genes carried by humans and covers the majority of drug targets. Our CRO service team possesses solid theoretical knowledge and extensive project service experience in clinical drug targets, providing comprehensive project support to expedite the drug screening process.

Full-length Coverage of Pathogenic Genes

Cyagen self-developed TurboKnockout^® technology and BAC fusion technology overcome the challenges of large-scale genomic segment replacement. This enables complex model gene editing, achieving full-length coverage of disease hotspot mutation regions (including exons, introns, and both UTR regions). HUGO-GT™ offers a higher degree of humanization compared to traditional models, meeting the project requirements of most pharmaceutical companies and researchers without patent or ownership disputes.

Personalized Customization

Based on the extensive project experience, the professional R&D team of Cyagen provides efficient whole genomic DNA humanized customization services for HUGO-GT™ model development. Not only can we customize site-specific targeting of large gene segments up to 350 Kb, but we could also perform gene re-modification on existing HUGO-GT™ wild-type humanized ES cells, which allows for rapid generation of disease models, shorter construction time, and higher positive rates, thereby providing experimental animal models as well as littermate controls for research purposes.

HUGO-GT™ Case Study: Ophthalmology-related
hRHO Mouse Model

The rhodopsin protein encoded by the human RHO gene is necessary for normal vision, and is critical for the transmission of low-light signals. Mutations in the RHO gene have been identified as associated with ophthalmic diseases such as retinitis pigmentosa (RP) and congenital stationary night blindness (CSNB). Currently, gene therapies targeting RHO, such as ASO and CRISPR, are being developed. The application of the HUGO-GT™ whole genomic DNA humanized animal model will help advance potential RHO-related therapies towards clinical trials.

Cyagen utilized embryonic stem (ES) cell gene editing and targeting technology to replace a 500bp fragment downstream of the Rho gene ATG promoter in C57BL/6J mice with a 500bp fragment downstream of the human RHO gene ATG promoter, thus creating the B6-hRHO mouse. Based on this, we further constructed the B6-hRHO (P23H) mouse model with a hot-spot mutation.

The results here indicated that the RHO protein encoded by the human gene was expressed normally in the B6-hRHO mouse, and its retinal morphology and function were similar to those of the wild-type mouse, showing no visual defects. However, the B6-hRHO (P23H) mutation mouse exhibited a phenotype of retinitis pigmentosa. These data demonstrated the successful construction of B6-hRHO and B6-hRHO (P23H) models, which can be used to study the pathogenic mechanisms of retinitis pigmentosa and the development of related new drugs.

No obvious alterations in the eye and retinal morphology of humanized hRHO mice while significant disruption of retinal structure could be observed after mutating P23H of hRHO.