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TurboKnockout-Pro全人源化小鼠 抗体药物研发、基因治疗重要研究工具 无专利风险 无脱靶效应 高同源重组效率
TurboKnockout-Pro for Whole Gene Humanization in Mice

An Important Research Tool for Gene Therapy & Antibody Drug Development

No Off-target Effects
Freedom to Operate
Large Fragment Knockin (up to 300kb)

The biopharmaceutical industry is in a critical period of innovation and transformation, with rapid developments due, in part, to evolving research tools and techniques. As an important research tool in the process of scientific research verification and preclinical drug development, genetically modified model animals can greatly improve the success rate of drug development and save a lot of translational research costs. However, the mouse biological system is not completely consistent with that of humans, especially shown in the immune system differences. Given that many human pathogenic factors and drugs are germline-specific, it is particularly important to establish an effective humanized mouse model for the pathogenic process studied.

TurboKnockout-Pro - Cyagen's Gene Editing Technology Helps Drug Development

TurboKnockout-Pro gene editing technology innovatively combines TurboKnockout® technology independently developed by Cyagen with BAC recombination technology for improved technological capabilities. Due to the limitation of the capacity of DNA cloning vectors, the exogenous gene fragments used to make genetically modified animals are usually less than 30kb, so some important elements that regulate gene activity are inevitably lost. However, BAC can accommodate foreign DNA up to 300kb. Through BAC recombination, longer fragments of genes and regulatory sequences can be introduced, so as to be closer to the expression pattern of endogenous genes and retain a greater degree of genetic humanization.

Compared with traditional ES cell gene targeting technology, TurboKnockout-Pro not only has the advantages of accurate modification and stable integration effect, but also can greatly shorten the project cycle by realizing multi-step BAC recombination at the ES level. Compared with CRISPR/Cas9 technology, TurboKnockout-Pro technology has no off-target effects and no patent disputes, providing freedom to operate. For these reasons, it is a commonly used technology for new drug research and development projects.


Application Type 1:
Generation of Whole-Gene Humanized Mice

TurboKnockout-Pro gene editing technology integrates BAC recombination to achieve large fragment gene knock-in of up to 300kb without off-target effects. This enables the introduction of more complete human genes into mouse cells to generate whole-gene humanized mouse models. The upgrade of technology saves project development costs by reducing the number of gene editing roundsand is free from patent disputes, which together accelerate the development of new drugs. Case Study: Humanized CD3 Mouse Model

The interspecies sequence preservation is relatively low in the extracellular domains of CD3ε (47% homology between human and mouse) at the amino acid level, CD3δ (57% homology between human and mouse), or CD3γ (60% homology between human and mouse). Therefore, therapeutic antibodies specific for human CD3 cannot effectively activate mouse effector T cells. The establishment of an experimental animal model suitable for evaluating human CD3-specific therapy is required. Cyagen has used TurboKnockout-Pro technology to successfully build a fully genetically humanized CD3 mouse model for an immunotherapy development company, helping the development of innovative multispecific antibody drugs. In addition, Cyagen has cooperated with a number of scientific research institutions, enterprises and institutions to build several large fragment humanized mouse models via TurboKnockout-Pro.


Application Type 2:
Generation of Whole-Gene Humanized Point Mutation mice

The expression of gene products seen in human disease is not always as high as a transgene, but it should still express to an extent, unlike the non-expression seen in a gene knockout. In many cases, the occurrence of genetic diseases is due to the change of protein structure caused by the mutation of single or several nucleotide bases, which is manifested as abnormal protein activation or inhibition. Therefore, constructing precise gene mutation models that are refined to characterize disease has become the best solution for effective preclinical and translational research. When the human gene under study has a high homology with the mouse gene homologous sequence, it can be simulated by directly making a point mutation mouse model. When the sequence homology between human and mouse is low, it is necessary to make humanized mice with mutations to better mimic human disease phenotypes. In the gene therapy research strategy, the causative gene(s) are found by genomics or proteomics, then combined with the humanized point mutation mouse model for verification, and this has led to many breakthrough discoveries and high-impact publications.

Construction Process of Fully Humanized Mouse Model
* Click the button to view the Turboknockout-Pro target vector design and construction strategy diagram
Advantages of Cyagen's Whole-Gene Humanized Mice Platform
TurboKnockout-Pro TurboKnockout® Traditional Gene Targeting CRISPR/Cas9 Gene Editing
Turnaround time 6~8 months 6~8 months 12-14 months 5-7 months
Species C57BL/6N, C57BL/6J, BALB/c C57BL/6N, C57BL/6J, BALB/c C57BL/6N, C57BL/6J, BALB/c No limits
Size of inserts ~ 300 kb ~20 kb ~20 kb ~15 kb
100% ESC-derived F0 Yes Yes No No
Potential off-targeteffects No No No Yes
Selection cassette self-deletion Yes Yes No N/A
IP Issue No No No Potential
No off-target effects, accurate modification, shorter turnaround
TurboKnockout-Pro gene editing technology is built on the traditional ES cell gene technology. It not only has the advantages of accurate modification and stable integration effect, but also generates 100% ESC-derived founder mice, rather than chimeras, and uses a self-deleting selection cassette to reduce two generations breeding time, providing founders in as fast as 6 months. At the same time, TurboKnockout® can perform multi-step BAC recombination at the ES level, which can further shorten the experimental period for complex genetic modifications.
Freedom to operate
Compared with CRISPR/Cas9 technology, TurboKnockout-Pro technology is free from patent disputes and is a commonly used technology for new drug research and development projects.
Professional technical support
Our professional project management team will balance speed, quality, and price according to your preferences or project needs, design the experimental plan for you attentively, and provide timely project reports and after-sales service.
Successful cases help new drug research and development
Using TurboKnockout-Pro, Cyagen has cooperated with a number of scientific research institutions, enterprises and institutions to build large fragment knockin (LFKI)humanized mouse models, such as the successful construction of whole-genetic humanization CD3 mouse models for specific antibody drug development.
Upcoming Event
Webinar: Genomically Humanized Mouse Models: Generation and Application in Biomedical Research

April 13, 2022 @ 22:00 pm PT
April 14, 2022 @ 14:00 pm JT

This webinar is presented by Cyagen’s Chief Scientific Officer, Dr. Marvin Ouyang, who has over 20 years’ of dedicated experience developing custom genetically engineered rodent models.

In this Webinar, you will learn:
  • What are TurboKnockout® and TurboKnockout® Pro
  • Genomically Humanized Mouse Model in GCT Drug Development
>> Register Now
About Cyagen

With over 15 years of experiences, Cyagen provides comprehensive in vitro and in vivo research model services for the basic and preclinical research stages of Gene & Cell Therapy (GCT) drug development. We can provide cell models and animal models that regulate the expression of target genes, as well as a series of related services, such as phenotyping, functional verification, pathological analysis and more. We have an experienced team of experts and a mature technology platform, with customers in dozens of countries and regions around the world. Our experts are ready to work with you to find the best plan for your goals.

>> Request a Free Consultation Now