We provide a service to generate transgenic mice by traditional pronuclear microinjection. Using this method, transgenes are inserted randomly into the genome.

Expression of the transgene and the subsequent transgenic mouse phenotype may vary from line to line due to the nature of random gene insertion. We recommend to generate multiple independent trangenic mouse lines in order to interpret the experimental results.

Available mouse strains: FVB, C57BL/6

We also offer Knock-in and Knock-out Mice and Knock-in and Knock-out Rat services. For more information please contact us.

Generation of transgenic mice by random insertion

Step 1: Cloning of cDNA, promoter of interest, and other elements

Step 2: Construction of the transgene vector

Step 3: Pronuclear microinjection of vector DNA into mouse embryos

Step 4: Transfer of injected embryo into recipient mothers

Step 5: Genotyping of offspring to identify potential transgenic founders

Step 6: Breeding to F1 or F2 progenies (optional)

We provide a high quality service for the generation of gene-targeted knock-in and knock-out mouse models. Our team has decades of experience in vector design, ES cell targeting and mouse handling. We will discuss your project needs with you and custom design your knockin/knockout mouse model. Our service includes consultation on available gene targeting vector design strategies and project assessment based on your model requirements.

Service Milestones for the Generation of Knock-in/Knock-out Mouse Models

• - Gene targeting vector construction and sequencing
• - ES cell targeting, screening and expansion of positive ES cell clones
• - Karyotyping of targeted ES cells
• - Cre/FLP recombination (optional)
• - Chimera production by blastocyst injection
• - Breeding of chimeras for germline transmission
• - Genotyping of offspring
• - Transfer of heterozygous targeted mice to the customer
• - Optional: breeding to generate homozygous mice

Features

Cost-effective

• - Targatt has years of experience and an exceptionally high success rate in generating conventional gene-targeted mice with our proprietary mESC lines.

High-Quality

• - Our scientists have extensive experience in gene targeting and genetic mouse models. Our services always include a rigorous quality control with multiple check-points to ensure your projects are completed with highest accuracy. All mice are generated in and shipped from our California facility (NIH guidance certified).

Please contact us with any questions regarding services not listed. We are happy to discuss your project with you and design customized strategies. We also offer Transgenic Mouse (Random Insertion) and Transgenic Rat Services (knockin, knockout).

Testimony of Dr. Zhenheng Guo, PhD, Assistant Professor of Internal Medicine, Division of Endocrinology and Molecular Medicine, University of Kentucky

Project: Germline-transmitted, conditional knock-out mice by tetraploid complementation

"Overall, I am very satisfied with the quality of your service. After we received the mice, we did extensive studies to characterize them. All of the PCR reactions confirm the gene modifications are in the correct sites. We have also crossed mice with Cre mice to demonstrate that the Cre deleted the exon in heterozygous pups as expected. All data obtained showed that the mice were correctly targeted. In addition, I am very happy with the frequent communication with your scientists during the process. I highly recommend your services."

Project types

Knock-in Mouse Models

The introduction of a gene into a specified location of the mouse genome can be used for various applications. Mice can be homozygous or heterozygous for the inserted gene.

• - Reporter genes (e.g. GFP, lacZ) are used for expression analysis of a gene of interest. The reporter gene is inserted into the gene of interested in-frame (non-disruptive), allowing for visualization of temporal and spatial gene expression pattern.
• - Humanized disease models can be generated by inserting a human mutant gene or gene fragment into the corresponding mouse gene. The diseased human allel is thus transcribed in the appropriate genomic context and can be analyzed on behavioral, pathological, cellular and/or molecular level.
• - DNA recombinases (CRE, FLP) can be inserted into a gene of interest. The celltype-specific expression of the recombinase then allows for gene inactivation in the desired tissue after crossing this knock-in mouse with a conditional knockout mouse.

Knock-out Mouse Models

The knockout technology is most commonly used for gene inactivation, either in a constitutive or conditional fashion.

• - Constitutive knock-out mouse models are widely used to study gene function. The gene of interest is permanently inactivated in all cells of the animal. However, this non-conditional knock-out may cause lethality and is therefore not always recommended.
• - Conditional knock-out models are inducible - the targeted gene is excised after crossing the mouse with a Cre-transgenic mouse line. Gene inactivation can be celltype-specific and/or chemically induced.

Gene knock-down using short hairpin RNA (shRNA) to inhibit expression of the corresponding gene has been widely used in gene function studies, drug discovery, and disease research.  Our TARGATT Technology allows us to generate in vivo shRNA knock-down mouse models in 3 months.

Targatt can create site-specific knock-in mice for you within 3 months. Using our novel TARGATT system, a gene of interest can be inserted at a well-characterized, transcriptionally-active locus in the mouse genome with guaranteed transgene expression. Tissue-specific and/or ubiquitous expression options are available. Please contact us for a list of plasmid construct with reporter genes. 

In addition to our full service we also offer individual service options to generate your TARGATT knock-in mouse.

You can prepare the DNA using the TARGATT Kit and we do the pronuclear microinjection for you. Or we can prepare the DNA that you then inject after setting up your own TARGATT mouse colony.

Please contact us to discuss your project plan for your fast & site-specific knock-in or TARGATT Knock-down Mouse services.