Next-Generation Humanized Mouse Models Overcome Traditional Limitations in Genetic Research

An exclusive interview with Dr. Yingbin (Marvin) Ouyang, Executive Vice President and Chief Scientific Officer at Cyagen, on breakthrough advancements in humanized genomic ortholog mouse models

Quick Overview:

  • HUGO mice leverage TurboKnockout® technology to achieve megabase-level gene replacement
  • HUGO-Ab™ models produce fully human antibodies with high affinity and low immunogenicity
  • HUGO-GT™ models feature whole-genome humanization for precise disease modeling & gene therapy research
  • AI integration accelerates model development and antibody discovery
  • Applications span rare diseases, ophthalmology, neurodegenerative disorders, and more

Breakthrough in Humanized Mouse Models: The HUGO Series

Cyagen has developed the next-generation Humanized Genomic Ortholog (HUGO) mouse models to overcome traditional  critical limitations in traditional genetic humanization approaches. These innovative HUGO humanized mouse models  leverage proprietary TurboKnockout® ES targeting technology to achieve megabase (Mb)-level in situ gene replacement, significantly shortening model development timelines while enhancing research accuracy. These advancements provide critical support for antibody drug discovery and gene therapy, enhancing efficiency in novel drug development and improving translational outcomes.

"With the rapid development of gene therapy and antibody development industries, there is increasing demand for highly precise humanized animal models," explains Dr. Yingbin (Marvin) Ouyang, Executive Vice President and Chief Scientific Officer at Cyagen. "Through the HUGO Program, we've addressed the fundamental shortcomings of traditional humanized gene mouse models, such as limitations in gene expression regulation, inadequate humanized regions, and complex model construction. By overcoming these obstacles, the HUGO series provides researchers with more precise tools for gene therapy and antibody development."

Meet Dr. Yingbin (Marvin) Ouyang: The Expert Behind HUGO

Yingbin (Marvin) Ouyang, MD, PhD

Executive Vice President and Chief Scientific Officer, Cyagen

Dr. Ouyang holds a Ph.D. from the Academy of Military Medical Sciences and previously served as a senior scientist at the Oklahoma Medical Research Foundation in the United States. With over 20 years of experience in genetic engineering research involving rodent models, he has successfully developed thousands of transgenic and knockout/knockin mouse and rat models. He has published numerous papers in high-impact academic journals such as PNAS and JBC. Furthermore, his technical services have been directly cited in hundreds of articles from top international journals, including Nature.

Limitations of Traditional Animal Models in Gene Therapy Research

Traditional genetically engineered humanized mouse models face several critical limitations that impact research accuracy:

  • Random gene insertion can disrupt mouse functional genes and generate unrelated phenotypes
  • Ectopic expression at non-endogenous loci unpredictably affects expression patterns
  • Multi-copy insertions alter gene dosage, affecting transgene expression levels
  • CDS-only insertions fail to replicate human gene expression accurately
  • Limited targeting scope prevents evaluation of intronic sequences, UTRs, and promoter regions

"Traditional transgenic methods randomly insert human genes into the mouse genome, which can lead to changes in the DNA sequence around the insertion site," Dr. Ouyang explains. "This can generate phenotypes unrelated to the transgenic sequence, thereby affecting the accuracy of the research results."

The HUGO-GT™ Solution: Whole-Genome Humanization

To address these limitations, Cyagen developed the HUGO-GT™ (Humanized Genomic Ortholog for Gene Therapy) model. By performing in situ replacement of mouse genes, these models offer significant advantages:

  1. Full-length gene in situ replacement retains regulatory sequences, allowing human gene expression at physiological levels
  2. Inclusion of human gene introns preserves the complexity of human splice isoforms
  3. Broader targeting capabilities for gene therapy evaluations targeting introns, UTRs, and non-coding sequences
  4. Extensive research applications across rare diseases, ophthalmology, neurodegenerative disorders, and metabolic diseases
  5. Faster construction and simpler breeding through TurboKnockout® technology
  6. International quality standards with production in AAALAC-accredited facilities

"The HUGO-GT™ model is better suited for simulating human diseases and provides a more efficient and reliable experimental tool for gene therapy research," notes Dr. Ouyang.

HUGO-Ab™: Revolutionizing Antibody Discovery

The HUGO-Ab™ fully human antibody mice represent another significant advancement in Cyagen's portfolio. These models carry complete sets of human immunoglobulin genes, enabling in vivo production of fully human antibodies with high affinity and low immunogenicity.

"The core advantage of the HUGO-Ab™ fully humanized antibody mice is that they can produce fully human antibodies with high affinity and low immunogenicity in vivo," Dr. Ouyang states.

The HUGO-Ab™ series includes:

  • HUGO-Mab™: Fully human monoclonal antibody mice
  • HUGO-Light™: Fully human common light chain antibody mice
  • HUGO-Nano™: Fully human nanobody mice

Five Core Advantages of HUGO-Ab™ Models

  1. Complete product range with multiple series and background strains
  2. Independent intellectual property rights with proprietary TurboKnockout® technology
  3. Comprehensive humanization of antibody heavy chain, Kappa, and Lambda light chain variable regions
  4. Enhanced immune response with HUGO-Ab-eKO™ variants for high-affinity antibody generation
  5. Flexible commercial model with one-time fee licensing or co-development options

Real-World Applications: Case Studies in Disease Research

The HUGO-GT™ models demonstrate exceptional utility across various disease research areas:

  • B6-hSMN2: For studying spinal muscular atrophy (SMA)
  • B6-htau series: For Alzheimer's disease (AD) and frontotemporal dementia (FTD) research
  • B6-hRHO and B6-hABCA4: For retinal diseases like retinitis pigmentosa (RP) and Stargardt disease

"Take the SMN2 fully humanized mouse model B6-hSMN2 of Spinal muscular atrophy (SMA) as an example," Dr. Ouyang shares. "We used it to test an antisense oligonucleotide (ASO) targeting SMN2. The results showed that ASO treatment significantly increased SMN protein expression in the brain and enhanced the number of motor neurons in the anterior horn of the spinal cord."

The positive outcomes included improved survival rates and delayed tissue pathology, validating the model's effectiveness in preclinical research.

Proprietary Technology: TurboKnockout® ES Targeting

The technological foundation of HUGO mouse models lies in Cyagen's proprietary TurboKnockout® ES targeting technology. This advanced approach enables:

  • Establishment of ES cell lines with superior genetic efficiency
  • Targeted microinjection at specific embryonic development stages
  • Complete replacement of endogenous ES cells without chimeric intermediates
  • Model generation in as little as four months
  • Self-deleting Neo Cassette system for marker-free heterozygous mice

"Through microinjection at a specific stage of embryonic development, TurboKnockout® ES cells can fully replace endogenous ES cells, effectively bypassing the traditional 'chimeric' stage," explains Dr. Ouyang. "This significantly shortens the timeline for ES cell-based targeting."

AI Integration: Enhancing Model Development and Applications

Cyagen has integrated artificial intelligence technology to strengthen its HUGO mouse model platforms:

Rare Disease Data Center (RDDC)

The AI-powered RDDC platform analyzes genomic data through machine learning algorithms to:

  • Identify potential pathogenic genes and regulatory elements
  • Enable precise gene in situ replacements
  • Guide customized mutations
  • Improve disease simulation accuracy
  • Support pathogenic mechanism research

AbSeek™ Antibody Intelligent Computing Platform

This platform combines AI with bioinformatics technologies to support:

  • Comprehensive antibody development stages
  • Sequence analysis and structure prediction
  • Lead compound optimization
  • Antibody molecule generation
  • Automated antibody design workflows

"When combined with our HUGO-Ab™ fully human antibody mice, the AI antibody platform accelerates the antibody discovery process, shortens development timelines, improves success rates, and provides full-cycle support for antibody research," Dr. Ouyang highlights.

Future Directions for HUGO Mouse Models

Cyagen plans to continue advancing its HUGO series models through:

  1. Expanded product lines covering additional disease areas
  2. Global research collaborations with leading institutions
  3. Enhanced customization services for specialized research needs
  4. Comprehensive CRO support to accelerate drug development

"We believe that the HUGO series mouse models will play an increasingly important role in future biomedical research," concludes Dr. Ouyang.

Interview with Dr. Marvin Ouyang: Key Insights on HUGO Series Development

Below are selected highlights from our in-depth conversation with Dr. Ouyang about Cyagen's HUGO series technology and applications.

Q: In recent years, significant progress has been made in gene therapy, with animal models playing a crucial role. Could you discuss the current limitations of animal models in gene therapy research?

Dr. Ouyang: Animal models, especially genetically engineered humanized mouse models, are indispensable in gene therapy research. However, traditional genetically engineered humanized mouse models have several limitations. First, traditional transgenic methods randomly insert human genes into the mouse genome, which can lead to changes in the DNA sequence around the insertion site, such as fragment deletions or disruption of mouse functional genes. This can generate phenotypes unrelated to the transgenic sequence, thereby affecting the accuracy of the research results.

Second, ectopic expression at non-endogenous loci can unpredictably affect expression patterns and disease mechanisms, with weak associations to human diseases. Moreover, multi-copy insertions can alter gene dosage, thus affecting transgene expression levels. While some humanized mouse models are constructed through targeted insertion, most insertions involve coding sequences (CDS), as full-length genomes are too large to allow for targeted insertion of the entire gene. This can result in human gene expression levels and spatiotemporal expression not aligning with the actual in vivo situation.

To address these issues, we have developed the HUGO-GTTM (Humanized Genomic Ortholog for Gene Therapy) model. By performing in situ replacement of mouse genes, we have successfully constructed a whole-genome humanized mouse model that covers a wider range of intervention targets. The HUGO-GTTM model is better suited for simulating human diseases and provides a more efficient and reliable experimental tool for gene therapy research. The specific advantages include:

  1. Full-length gene in situ replacement that retains regulatory sequences, allowing human gene expression to closely align with physiological levels.
  2. Inclusion of human gene introns, preserving the complexity of human splice isoforms, which is beneficial for studying the regulation of human gene expression.
  3. More suitable for modern gene therapy research, especially for gene therapy evaluations targeting introns, UTRs, and other non-coding sequences.
  4. Broader research applications, covering various disease research fields such as rare diseases & genetic disorders, ophthalmic diseases, neurodegenerative diseases, metabolic diseases, etc.
  5. Utilization of the unique TurboKnockout® technology, which enables faster construction, and due to the targeted insertion, breeding and maintenance are simpler compared to traditional knockout plus transgenic models.
  6. Strict quality control: All animals are produced in AAALAC-accredited and OLAW-recognized facilities, meeting international standards.


Q: Could you briefly introduce which models are included in Cyagen's HUGO Program?

Dr. Ouyang: Cyagen's HUGO series mice are a significant achievement in the field of genetically engineered humanized mouse models. It includes two major series: HUGO-Ab™ fully human antibody mice and HUGO-GT™ whole-genome humanized mice. These models are developed based on our proprietary TurboKnockout® gene editing technology, which allows for better simulation of human physiological and pathological characteristics, providing powerful tools for drug development and disease research.

For example, the HUGO-Ab™ fully human antibody mice carry the complete set of human immunoglobulin genes, enabling them to produce fully human antibodies with high affinity and low immunogenicity in vivo. The HUGO-GT™ whole-genome humanized mice are equipped with a more efficient large fragment vector fusion technology, which can serve as a universal template for targeted mutation customization services. Additionally, based on the HUGO-GT™ mice, we also offer CRO services for disease research fields such as ophthalmology, neurology, and tumor immunology, fully empowering research on genetic diseases and gene therapy drug development.

Q: What are the core advantages of the HUGO-Ab™ fully human antibody mice?

Dr. Ouyang: The core advantage of the HUGO-Ab™ fully humanized antibody mice is that they can produce fully human antibodies with high affinity and low immunogenicity in vivo, which can be summarized by five "full" attributes.

First, we have a full range of humanized antibody mouse products. We offer multiple product series, such as HUGO-Mab™ fully human monoclonal antibody mice, HUGO-Light™ fully human common light chain antibody mice, and HUGO-Nano™ fully human nanobody mice. The antibody heavy chain and light chain variable region genes of these mice are all human-derived, offering a higher degree of humanization and being closer to the true diversity of human antibodies.

Second, we have complete independent intellectual property rights. Our self-developed TurboKnockout® ES targeting technology enables Mb-level gene in situ replacement for knock-in and knockout, with no off-target effects. The gene modification is precise, stable, and the intellectual property rights are clear, making it the preferred gene editing technology for new drug development projects.

Third, fully humanized antibody development. The antibody heavy chain, Kappa, and Lambda light chain variable region genes are human-derived, offering more comprehensive humanization. Humanization of the Lambda light chain variable region germline genes enhances antibody diversity, making it closer to the true diversity of human antibodies.

Fourth, all models have high immune response capabilities. Based on the HUGO-AbTM fully human antibody mice, the "HUGO-Ab-eKO™ mice" are designed to knock out mouse genes for highly homologous targets, allowing the mice to generate a stronger immune response. This enhances the success rate of antibody screening and makes it easier to obtain high-affinity, highly specific, and cross-reactive antibody molecules.

Q: What is the key breakthrough that allows the TurboKnockout® technology to achieve Mb-level gene in situ replacement? Compared to other gene editing technologies, what are its unique advantages in terms of precision and stability?

Dr. Ouyang: The key breakthrough of the TurboKnockout® technology in achieving Mb-level gene in situ replacement lies in its unique model construction and gene modification techniques. This technology establishes TurboKnockout® embryonic stem (ES) cell lines with superior genetic efficiency. Through microinjection at a specific stage of embryonic development, TurboKnockout® ES cells can fully replace endogenous ES cells, effectively bypassing the traditional "chimeric" stage. This significantly shortens the timeline for ES cell-based targeting, with model generation achievable in as little as four months. Additionally, TurboKnockout® technology uses a unique Self-deleting Neo Cassette, which ensures that when the resulting mice are crossed with any strain, the Neo cassette is 100% self-deleted. This enables the rapid generation of heterozygous mice free of the selection marker.

Q: The HUGO-GTTM mouse model has a wide range of applications in disease research, right? Could you provide us with a few example cases?

Dr. Ouyang: Yes, the HUGO-GTTM mouse model has a very wide range of applications. For example, we have the B6-hSMN2 mouse model for studying spinal muscular atrophy (SMA); the B6-htau series mouse model for Alzheimer's disease (AD) and frontotemporal dementia (FTD) research; and the B6-hRHO and B6-hABCA4 mouse models for studying retinal diseases such as retinitis pigmentosa (RP) and Stargardt disease. These models can accurately simulate the pathological features of human diseases in mice, providing important tools for studying disease mechanisms and developing drugs.


Take the SMN2 fully humanized mouse model B6-hSMN2 (SMA) as an example. We used it to test an antisense oligonucleotide (ASO) targeting SMN2. The results showed that ASO treatment significantly increased SMN protein expression in the brain and enhanced the number of motor neurons in the anterior horn of the spinal cord. More importantly, the survival rate of the ASO-treated mice was markedly improved, and tissue pathology was delayed. This demonstrates that the HUGO-GTTM mouse model can effectively reflect drug efficacy in testing and provides reliable data support for preclinical research.

Q: What are Cyagen’s future plans for the HUGO series mouse models?

Dr. Ouyang: In the future, Cyagen will continue to deepen the research, development, and application of the HUGO series mouse models. On one hand, we will further expand the product lines of the HUGO-AbTM and HUGO-GTTM mouse models by developing more models that cover a wider range of disease areas. On the other hand, we will strengthen collaboration with researchers around the world, offering higher-quality customized services and CRO support to accelerate the development of innovative drugs and the study of disease mechanisms. We believe that the HUGO series mouse models will play an increasingly important role in future biomedical research.

Q: As AI technology is increasingly applied in the biomedical field, has Cyagen's HUGO series mouse models also integrated AI technology? Could you share how AI is empowering the HUGO-GTTM and HUGO-AbTM mouse models?

Dr. Ouyang: At Cyagen, we are committed to integrating cutting-edge technologies into our R&D efforts—and AI is no exception. We have leveraged AI technology to provide stronger support for the development and application of both HUGO-GTTM and HUGO-AbTM mouse models.


We utilize our AI-powered Rare Disease Data Center (RDDC) platform to analyze and mine vast amounts of genomic data. Through machine learning algorithms, the RDDC platform can rapidly identify potential pathogenic genes and regulatory elements, enabling us to perform more precise gene in situ replacements and customized mutations. This not only improves the efficiency of model construction but also enhances the accuracy of disease simulation in humans. By leveraging extensive bioinformatics data, we conduct in-depth research on pathogenic mechanisms and have developed a series of HUGO-GTTM whole-genome humanized mouse models targeting specific genes—empowering genetic disease research and gene therapy drug development.


In the field of antibody discovery and antibody engineering, we have developed the AbSeek™ Antibody Intelligent Computing Platform, which integrates cutting-edge artificial intelligence and bioinformatics technologies. Equipped with both proprietary and open-source computational and visualization modules, the platform offers outstanding speed and accuracy. The AbSeek™ platform comprehensively supports all stages of antibody development—from screening and optimization to validation—providing a wide range of tools including antibody sequence analysis, structure and function prediction, lead compound optimization, and antibody molecule generation. This enables a simplified and automated antibody design workflow, driving efficient and innovative progress in antibody engineering. When combined with our HUGO-AbTM fully human antibody mice, the AI antibody platform accelerates the antibody discovery process, shortens development timelines, improves success rates, and provides full-cycle support for antibody research.

About Cyagen

Founded in 2006, Cyagen is a global provider of genetically modified rodent models and innovative cell and gene therapy solutions for research and development. The company has established extensive collaborations with scientists and institutions in over 100 countries, contributing to more than 10,925 academic publications, including articles in the prestigious journals Cell, Nature, and Science.

From animal model development to AI-powered tools for data analysis and therapeutic discoveries, Cyagen provides comprehensive solutions for accelerating basic research and new drug R&D through its unique offering of models, data, algorithms, and services.

For more information about HUGO series mouse models and Cyagen's services, visit www.cyagen.com

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