カスタムのノックアウトマウスモデルの作製と維持

遺伝子改変ノックアウトマウスは、マウスゲノムを改変できるという特性のため、ヒト疾患をモデル化するための強力なツールとなっています。Charles Riverでは、数千ものノックアウトマウスモデルで世界中の数百のお客様をサポートし、研究準備の整ったコホートを提供してリサーチのニーズを満たしています。

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トランスジェニックマウスとラットのモデルの作製

ノックアウトマウスを作製するためのゲノム編集ゲノム編集のためのCRISPR-Cas9*、動物モデル作製、遺伝子治療、およびヒト疾患のモデリングの詳細については、ウェビナーシリーズをご視聴ください。
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Charles Riverの科学チームは、機能ゲノミクスおよび分子工学の分野のリーダーである業界パートナーと協力関係を結んでいます。インビトロ およびインビボ の専門知識を結集し、マウスの作製、特性化、保存、提供に最適な環境を整えます。


Charles Riverが提供するマウスモデル関連サービス

 

Types of Knockout Mouse Models

  • Constitutive Knockout Mice

    This model is engineered to carry an inoperative gene. Usually, inactivation of the gene is achieved by the deletion (partial or total) of its sequence, and it is nonfunctional in the entire animal.

  • Conditional Knockout Mice

    These knockout mice allow the deletion of a gene in a tissue- and/or time-specific manner. Often, the conditional KO mouse is achieved through the Cre-lox system. Instead of deleting the critical sequence, it is flanked by loxP sites (termed a floxed sequence). Cre recombinase deletes the sequence between two loxP sites. An inducible or tissue-specific Cre is used to knockout gene function in only that tissue.

  • RNAi Knockout Mice

    RNA interference can reduce the expression of a target gene, without deleting it. Typically this is combined with conditional and/or inducible approaches to reduce expression of the target only under certain conditions in adult animals.


VIDEO: Considerations for Creating Knockout Mice and Other Transgenic Animal Models
 

  • Video Transcript

     


    00:08
    Using genetically engineered rodents as models of human disease, have transformed scientific research. Properly designed and validated, they can provide accurate study data, decrease the number of animals used and reduce study costs. Those working with mutant and transgenic mouse models often fall into two categories, academic research institutions looking to decipher the mammalian genome to understand how a specific gene functions and global pharma and biotech companies utilizing models such as knockout mice, in order to validate new drugs and treatments.
     
    00:46 Research employing these models continues to evolve at an incredible pace. How you obtain and maintain them is important because many factors can impact experimental reproducibility and scientific conclusions. The quickest method is to purchase them from a repository or obtain them from a collaborating lab. However, one must consider if these models are an exact fit for your research needs. What is their health status? Could their genetic integrity have been compromised? Could they be carrying the wrong genes?
    01:18 Many institutions have set up internal transgenic cores for generating new mouse models. However, they can sometimes be impacted by health status or capacity issues, which can influence project timelines. In addition, institutions may lack the expertise required for specific projects. CRISPR-Cas9 technology has accelerated the creation of knockout mice and other mutant animals. It not only reduces timelines but also allows for flexibility to work with other species such as the rat model, which may be a better fit for certain studies.
    01:53 But is CRISPR always the best choice? To answer this question, you should consider how complex the gene modification is. How to detect unexpected CRISPR off and on target effects. For commercial organizations, issues with IP and licensing may result in serious financial ramifications. An effective way to obtain your knockout, knockin or transgenic mice is to work with a qualified external vendor.
    02:20 You'll receive advice from experienced scientific teams, utilizing a variety of techniques, strict adherence to project timelines, access to and advice on CRISPR licensing with ownership of intellectual property, a high health status with breeding under VAF+ or SOPF standards and a high degree of model validation and analysis to ensure your new model generates consistent study data.
    02:46 Charles River's model creation services provide an optimal environment for generating, characterizing, preserving and distributing your transgenic lines. Our full service portfolio includes custom breeding, embryology services, genetic testing, drug discovery and safety assessment services. With vast experience in the drug development life cycle, you'll have a trusted partner to get you from concept to clinic. Learn how we can help by talking to someone from our scientific team today.

     

     

 

Frequently Asked Questions (FAQs) About Knockout Mice

  • What are some of the advantages of using knockout mouse models?

    The gene knockout method is the simplest approach to reveal fundamental gene functions. Conditional knockout/inductible mice, inducible knockout mice, and knockdown mice can avoid the risk of a lethal phenotype associated with gene inactivation at earlier stages in development and allow for an analysis of gene inactivation effects on adult animals. Furthermore, as 65% of protein coding genes are likely pleiotropic, a conditional knockout mouse simplifies phenotyping analysis by focusing on a specific cell type.

  • Are there other project considerations?

    Disrupting genes can sometimes result in compensation by other members of a multigene family. In these cases, multiple knockouts may be needed to obtain phenotypical effects (e.g., Hox genes). Moreover, it is possible that gene knockouts may fail to produce observable phenotypes in knockout mice, or may produce different characteristics from those observed in humans. Mouse background choice is an important consideration for better mimicking human phenotypes – ask our experts for further advice.

  • What are the methods used for generating a knockout mouse model?

    Depending on your study and objectives, knockout mice can be obtained through:


    For further information on knockout mice, please see the IKMC (International Knockout Mouse Consortium) website.

  • Where can I get more information about CRISPR?

    The Science of Controlling CRISPR
    With technologies like CRISPR gene editing accelerating at a rapid pace, what was once thought to be impossible to achieve with mice may become reality sooner than we ever imagined. The challenge will be to fully understand and control these new mouse model creation tools. For example: how does one limit off-target effects? What do we know about circular permutation, or inhibiting CRISPR with anti-CRISPRs? Read the full story

    The CRISPR Potential in Accelerating Drug Development
    CRISPR's comparative ease of use is expected to contribute to the development of more multifaceted cellular assays with improved predictability for drug therapies. Some of the benefits include reduced attrition rates of compounds and improved target validation. Read the full story

Rodent model genetic quality control guidebook for genetic monitoring

New Guidebook: Rodent Model Genetic Quality Control

Managing a genetically engineered rodent colony is not so simple. Get our new guidebook to obtain sound advice on colony management, genetic monitoring and embryology techniques required to protect your research.

Download Now

 

The Impact of Genetic Background and Genotyping Webinar: Questions and Answers

 

*CRISPR/CAS9 used under licenses to granted and pending US and international patents from The Broad Institute and ERS Genomics Limited.