Animal Models

Applied StemCell’s PrimeCKO™ Conditional Knockout (CKO) Mouse Models service will enable you to have precise control over where or when your gene of interest in knocked out. Using CRISPR and the Cre-Lox system, we can generate floxed Conditional Knockout Mouse Models for a variety of temporal and spatial knock-out conditions in mice. You can breed the “floxed” PrimeCKO™ conditional knockout mice with commercially available Cre mouse lines, or we can generate a custom Cre mouse model for high-level Cre expression.

• Affordable! Low prices and founder guarantee
• F1 breeding for germline transmission
• Novel CRISPR strategy for higher efficiency and success rate
• Animal IP belongs to customers
• Fast turnaround of 5-8 months

Our comprehensive service includes vector construction, pronuclear injection, and embryo transfer. The offspring are genotyped for integration, and we deliver at least 2 founders (or germ transmitted F1s). Schedule your free consultation today to discuss your conditional knockout mouse project with one of our experts.

Summer Mouse Sale! Conditional Knockout Mouse Model Generation Only $22K USD

Summer Sale! Simple Knockout Mouse Models Only $10K USD

ASC’s proprietary TARGATT™ knockin technology enables highly efficient and site-specific gene knockin mouse model generation. This technology uses serine integrase, PhiC31 (ΦC31) to insert any gene of interest into a preselected intergenic and transcriptionally active genomic locus that has been engineered with a docking site for stable, site-specific transgene integration.

• High-efficiency insertion (up to 65%)
• Site-specific into a genetically active safe harbor locus
• Large fragment knock-in (up to 22 kb)
• Single copy in an active locus: avoid gene silencing and genomic instability.
• High level, stable gene expression
• Germline transmitted F1 mice in 5-8 months

Mouse models: Gene overexpression, conditional expression, humanized gene knockin, gene knockdown, reporter genes

TARGATT™ Fast & Site-Specific Knock-in Mouse Service Workflow

Generate your gene knockout mouse model for inactivating genes either by frameshift mutation or by fragment excision, using CRISPR/Cas9. Gene knockout or disruption in mouse models provide an in vivo environment to study the function or role of the gene of interest. Applied StemCell has optimized and most up-to-date gene targeting strategies to generate the knockout mouse model of your choice. We can generate constitutive knockout mice or for a more advanced study of the gene function, conditional/ inducible knockout models in as little as 5 months (F1 germline transmitted).

Summer Sale! Simple Knockout Mouse Models Only $10K USD

Summer Mouse Sale! Conditional Knockout Mouse Model Generation Only $22K USD

Our affordable, custom knock-in mouse model generation service using two complementary technologies, CRISPR/Cas9 and TARGATT™, allows us to offer a diverse portfolio of knock-in modifications, gene-specific knockin (up to 6 kb) or safe harbor locus knockin (up to 10kb with CRISPR and up to 22 kb with TARGATT™). ASC's comprehensive knock-in service allows for the insertion of humanized genes, reporter genes, tags, and more. We work with you to design a specific project outline that captures all of your mouse knock-in model requirements.

• Locus specific (or) safe harbor locus knock-in (e.g., Rosa26, H11, others)
• Knock-in reporter genes/tags, human gene/exons, fusion genes, replacement, and more
  • Safe harbor knock-in: CRISPR (Rosa26, H11, AAVS1) or TARGATT™ (Rosa26, H11 or a custom locus).
• Efficient CRISPR strategies and protocols for large fragment knock-in
• F1 breeding for germline transmission
• Novel CRISPR strategy for higher efficiency and success rate
• Animal generated in an AAALAC-accredited animal facility
• Animal IP belongs to customers
• Fast turnaround of 5-8 months

At ASC, we utilize CRISPR/Cas9 technology to produce genetically modified humanized mouse models. Our experts can design your ideal project to knock in or replace your specific mouse gene with a human gene, genomic sequence, or regulatory element. We can even design humanized models with conditional gene activation. These animal models are ideal for drug development, efficacy and safety studies, and more.

• Most up-to-date CRISPR design strategies and protocols
• 100% target-site cutting efficiency using validated gRNA
• >99% success rate in generating final mouse model
• Wide variety of genome modifications are available
• F1 breeding to confirm germline transmission
• Electrophysiology and behavioral assessments for your custom mouse models

ASC also supplies an assortment of well-characterized, off-the-shelf humanized mouse models. If you would like to learn, contact us today to set up your free consultation.

Service to generate transgenic mice using BAC or Random insertion.

Humanized ACE2 Mouse Models Available Now

To aid the global research efforts to find a solution to the COVID-19 pandemic, Applied StemCell has leveraged its site-specific TARGATT™ knockin technology to engineer two fully humanized ACE2 (hACE2) mouse models with lung epithelium-specific expression driven by the K18 promoter. These TARGATT™ hACE2 mouse models have the K18-hACE2 transgene integrated into a preselected, well-characterized safe harbor locus (genomic hotspots): mH11 or mRosa26 and might be more representative to model the severity of the SARS-CoV-2 infection. The TARGATT™ hACE2 mouse models are ideal to help understand the severe pathogenesis of COVID-19, and to evaluate the efficacy of antiviral drugs and vaccines.

Worldwide shipping! 

Applied StemCell’s off-shelf mouse catalog offers >100 humanized immune checkpoint mouse models, with single, double, and triple humanized gene models.

• Knock-in of humanized counterparts of immune checkpoint gene into the endogenous locus.
• Proprietary human transgene knock-in strategy to enable screening of many different antibodies and immuno-therapy compounds.
• In vivo and in vitro validated for physiological expression of human version of the genes (in-house and through collaborators).
• Fully functional murine immune system  
• The humanized immune checkpoint mice have been generated in the most widely used C57BL/6J background used in immunology and oncology studies.
• Ideal for immunology and antibody/ immuno-oncology therapy efficacy studies and drug discovery research.

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Antibody Efficacy Testing Service

Leverage ASCs cost-effective evaluation of in vivo efficacy and fast timelines to screen your early-stage lead antibodies using our antibody efficacy testing platform:

Workflow includes:

• Project Proposal: details on the test antibody (and positive and negative control), project design, scope of the project
• Receive antibody(ies) from client
• Tumor cell implantation and development in humanized immune checkpoint mouse models (mean tumor size reaches 100-150 mm^3)
• Administration of antibodies to humanized immune checkpoint mouse tumor models
• In vivo evaluation of antibody efficacy: tumor size and body weight, twice a week for 3-4 weeks
• Final report with data and figures
  • Report will include comprehensive information about experimental/ project outline, project schedule, survey notes, raw data for efficacy parameters measured/ endpoints, calculated/ integrated results, raw and integrated figures (graphs and other figures corresponding to endpoint measurements).

* Endpoint: Tumor burden should not exceed 10% of the animal’s bodyweight. The study will be terminated with euthanizing all animals when the mean tumor volume reaches about 3000 mm^3.

Optional post-mortem tests include: Blood RT/Biochemical analysis; FACS analysis on PBMC (including CBA); serum/ plasma preparation and then ELISA, MSD or WB analysis, qPCR,WB, IHC, IF, TILs analysis

Timeline: 2-3 months after receipt of antibodies

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The Cre-Lox system is a powerful gene manipulation tool that enables controlled gene expression in a specific tissue or by induction in mouse models. Such controlled/ conditional gene expression or ablation enables deeper study of gene function without problems such as artificial phenotype(s) and embryonic lethality.

Applied StemCell offers several ready-to-use, mouse models expressing Cre recombinase under the control of various tissue-specific and/or inducible promoters as well as Cre-reporter mice:

• Well-characterized Cre driver mouse lines
• Generate temporal and/or spatial control of gene expression mouse models
• Breed with your loxP floxed mice or we can generate a custom floxed mouse for you using our PrimeCKo™ Conditional Mouse Model Generation Service; in your facility or ours

Available Cre Mouse Lines

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Cytokines are cell-to-cell signaling proteins that play a central role in immune development and responses, and its dysregulation is associated with a several diseases such as autoimmune and inflammatory disorders, chronic infections and even cancer. Applied StemCell offers research-ready, mouse models expressing humanized cytokine genes for studying the underlying mechanisms of immune response and inflammation as well as for immunotherapy and oncology efficacy studies and drug screening.

• Knock-in of humanized counterparts of cytokine gene(s) into the endogenous locus.
• In vivo and in vitro validated for physiological expression of human version of the genes (in-house and through collaborators).
• Mouse models generated in the widely used C57BL/6J background.

Cytokine Mouse Models

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With 11+ years of experience in genetically engineering mouse models (GEMM), Applied StemCell offers a wide-range of ready-to-use, constitutive knockout (KO) and conditional knockout (CKO) mouse models targeting various genes, with some very unique models available only here. Our GEMM catalog also includes reporter/ tag knock-in, inducible expression mouse models and other tool mice, to expand the scope of your research and screening applications.

• Genetically validated models
• Majority of the lines are in the C57BL/6 background
• Facilities operated under the strict AAALAC, SPF and IACUC guidelines
• Can be shipped worldwide
• Valuable research tools for genetics research, disease modeling and drug screening

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Applied StemCell provides next-generation immunodeficient mouse models such as our proprietary M-NSG (NOD-scid: non-obese diabetic severe combined immune deficiency, gamma), RAG1, RAG2 knockout (KO) models and more. Our M-NSG mice lack mature T cells, B cells, and functional NK cells.

• Highest degree of immunodeficiency
• Long-lifespan
• Excellent tumor xenograft (PDX & CDX) models with minimal immune rejection
• Ideal recipient for human immune reconstitution (humanized mice). Ex. hPBMC/ hHSC engraftment
• Applications include immuno-oncology, immunodeficiency, inflammation, transplantation and immune mechanism

Are you looking to further your preclinical studies and begin your in vivo efficacy, safety, and toxicity experiments? Applied StemCell now supplies research-ready humanized PBMC and CD34+ reconstruction mouse models ideal for infectious disease, gene therapy, hematopoiesis, drug target, and immunogenicity studies. Our humanized models are generated using our M-NSG immunodeficient mice through the injection of healthy human donor peripheral blood mononuclear cells (PBMCs) or CD34+ hematopoietic stem cells (HSCs). After a few weeks of reconstruction following the injection of PBMCs or CD34+ HSCs, the mice develop a human-like immune system and are ready to be shipped. Place your order today and claim your free t-shirt!

• Affordable
• Created in the U.S.A.
• Worldwide shipping
• In vivo efficacy, safety, and toxicity testing services with PBMC or CD34 humanized mice are available – Inquire or visit our in vivo assay page

Customizable Services! If you would like our team to produce a PBMC or CD34 humanized mouse model with your PBMCs or CD34+ HSCs, we can inject your cells into our NSG mice to produce the model you require for your research. If you need PBMCs, CD34+ HSCs, or immunodeficient mice for your own lab, we also provide tissue collection/cell isolation services, off-the-shelf cell products, and M-NSG mice.  Contact us today to learn more.

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Human Immune System Reconstruction Mouse Model Generation

The TARGATT™ plasmids are required to generate the donor plasmid that will be used with our TARGATT™ Transgenic kits and TARGATT™ “attP” mice for site-specific transgene integration in mice. Applied StemCell offers a number of plasmids with a unique combination of promoters and multiple cloning site to meet your special needs, and to ensure robust transgene expression:

• TARGATT™ 2 (CAG + Poly A)

• TARGATT™ 3 (no promoter + MCS)
• TARGATT™ 6.1 (CAG-L4SL-MCS-PolyA)
• TARGATT™ 7 (PGK-MCS-PolyA)
• TARGATT™ 8 (PCA-MCS-PolyA)
• TARGATT™ 9.1 (PCA-L4SL-MCS-PolyA)

Make your own TARGATT™ site-specific knock-in mouse model for inducible gene expression, gene overexpression, reporter gene insertion, knock-down by siRNA, humanized models and other applications.

New product options for the TARGATT™ “attP” mice! The TARGATT™ “attP” (H11) mice contain an “attP” docking site at the well-defined mHipp11 (H11) safe harbor locus. These mice are to be used with the associated TARGATT™ plasmids and TARGATT™ transgenic kit for site-specific, stable integration of your gene-of-interest into the preselected locus, with high insertion efficiency, a fast turnaround, and high level expression of the transgene.

The mice are available as: (1) Live animal breeding pairs; (2) Live animals; male (3-6 months old); (3) Cryopreserved embryos.

The TARGATT™ “attP” (H11) mice is a valuable research tool to generate site-specific, knock-in mice for applications such as gene overexpression, reporter gene insertion, knock-down by siRNA, inducible gene expression and more.

New! Upgraded Transgenic Kit (Version 3) and Protocols! The TARGATT™ Transgenic Kits is an essential component for the generation of TARGATT™ site-specific, knock-in transgenic mice. The kits are designed using the same highly optimized protocols that are used in our popular and successful Custom TARGATT™ Mouse Model Generation Service. The kits contain PhiC31 integrase mRNA, control mRNA and embryo-qualified miTE buffer, and are intended for the microinjection of a TARGATT™ donor plasmid (purchased separately) into the pronuclei of TARGATT™ “attP” mouse embryos.

The TARGATT™ Transgenic Kits offers an affordable, efficient and faster alternative for experienced scientists and transgenic core facilities to generate numerous site-specific knock-in mouse models in their own vivarium. 

The TARGATT™ Mouse Genotyping Kit allows for convenient genotyping of TARGATT™ Transgenic Mice via tail biopsy. The kit contains reagents, PCR primer sets, buffer along with detailed protocols and anticipated results. This kit is intended for use after TARGATT™ Transgenic mouse model generation using the TARGATT™ Transgenic Kit, TARGATT™ Plasmids, and TARGATT™ “attP” Mice to confirm the transgene integration into the TARGATT™ attP docking site at either the mHipp11 (mH11) or mRosa26 locus.

Applied StemCell can generate custom conditional knockout rat models using CRISPR/Cas9 or other suitable gene-editing techniques to insert the 5’ and 3’ flanking LoxP sequences into the endogenous locus of your gene of interest. These conditional knockout rats can be bred with commercially available Cre expressing rats or custom-engineered Cre expressing rats to generate your rat model with precise control over where and when your gene is knocked out.

ASC offers Bacterial Artificial Chromosomes (BACs) to generate rat models. BACs are ultra-low copy vectors that hold up to 300 kb of genomic fragments.

• High-quality service
• F1 breeding for germline transmission
• Customized projects for in vivo assessments (automated behavior/ locomotor activity, EEG/ ECG, and pharmacokinetics) as well as in vitro evaluations (electrophysiology, immunohistochemistry & other biochemical assays

Applied StemCell offers Knockout Mouse Model Generation for engineering constitutive gene deletion or mutation (frameshift) to render a gene of interest non-functional. Applied StemCell offers high-quality custom genetically engineered mouse model (GEMM) generation services operated out of vivarium in AAALAC-accredited facilities with strict IACUC oversight. Our knockout mouse model protocols are very optimized and efficient to offer a >97% success rate. Order projects with us a stress-free research animal model generation that will suit your basic research and drug development needs.

Key Features:

• Up-to-date CRISPR design strategies
• 100% target-site cutting efficiency using validated gRNA
• Highly successful at generating final rat model
• Wide variety of genome modifications are available
• F1 breeding to confirm germline transmission
• Animal IP belongs to researchers
• Electrophysiology and behavioral assessments for your rat models

With the need for physiologically relevant animals models on the rise for more predictive research findings, genetically engineered rat models are becoming increasingly popular. Get ahead of your competition by leveraging Applied StemCell’s expertise in CRISPR/Cas9 genome editing technology and animal model engineering to generate the ideal knockout rat model to meet your specifications and budget.

• Global leader in genetically engineered rat models (Nature article, “CRISPR mouse model boom, rat model renaissance”)
• Detailed discussion to understand project requirements, optimal project design for precise genetic modification, and dedicated project management for timely updates

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Rat models are very important models for specialized research where mouse models are not quite representative of the human physiology or behavior. Like mouse models, targeted genetic engineering in rat models has been revolutionized with CRISPR/Cas9 and other technologies (TARGATT™, TALENs and ZFNs). Applied StemCell offers knockin rat model generation using two technologies: CRISPR/Cas9 and TARGATT™ to enable the engineering a wide range of knockin modifications.

• Highly optimized and efficient protocols
• High-efficiency gRNA for high Cas9 target cutting efficiency
• In vivo gRNA validation 
• F1 breeding to confirm germline transmission
• Dedicated project management and timely milestone and final reports

Point mutation mouse models are generated by inserting or replacing in one or more variant nucleotides in selected gene(s). These models are very important to study the importance of specific nucleotides or amino acids in gene expression or protein function and for disease modeling. As one of the earliest providers of CRISPR-based custom mouse model generation services, Applied StemCell can generate defined constitutive or conditional point mutation knock-in models using CRISPR/Cas9. We can generate your point mutation founders/ F1 germline transmitted animals using our optimized and efficient protocols, with a high success rate. All mouse models are generated in our facilities under AAALAC accreditation and IACUC oversight. We can also ship animals worldwide or as cryopreserved embryos.

• Highly optimized and efficient protocols
• High-efficiency gRNA for high Cas9 target cutting efficiency
• In vivo gRNA validation 
• F1 breeding to confirm germline transmission
• Dedicated project management and timely milestone and final reports

Bacterial Artificial Chromosomes (BACs) are ultra-low copy vectors that can hold up to 300 kb of genomic fragments, making them ideal vectors for the introduction of entire genes including the regulatory regions into the rat genome. BAC transgenic rats are similar to other knock-in models in the expression pattern of the gene of interest, except that the BAC is randomly integrated into the genome as a transgene. Applied StemCell can create BAC knock-in and conditional knock-in rat models for you.

Our services include:

• BAC design and cloning strategy
• Cloning and sequence validation of the BAC tailor-made to your specifications
• Pronuclear microinjection of BAC into rat embryos
• Genotyping

Alternatively, you can choose to have us do the BAC modification strategy and cloning, and you can do the pronuclear injections and transgenic rat screening yourself.

Applied StemCell’s proprietary TARGATT™ technology enables generation of physiologically relevant transgenic rat models suitable for a variety of applications including reporter gene expression, gene knock-down, conditional gene expression and disease models. This technology uses the Phic31 integrase to mediate an irreversible integration of large transgene(s) into a preselected, safe harbor locus with very high efficiency.

• Site-specific, single copy transgene integration overcomes challenges associated with random integration
• TARGATT™ knock-in rats in 6-9 months
• Direct microinjection of the TARGATT™ reagents into rat zygotes

Using the TARGATT™ technology, ASC has also developed Neural Specific Cre-Rat Lines in a Sprague Dawley rat background.