Targeted Transgenesis: Rosa26 or Hprt Quick Knockin

Today's fastest, most robust, and financially affordable approach to develop a genetically modified rodent model is to insert your transgene into gene loci such as Rosa26 or Hprt (targeted transgenesis).

Strength of this approach:
  • Controlled gene insertion
  • 7 months' development time (heterozygous animals)
  • Line stability over many years
  • Delivers physiologically relevant and reliable results

Learn more and watch the video ➟

 

 

Strong Experience in Targeting Rosa26 and Hprt

  • "With the idea to develop a mouse model for the rare human disease ALD, we reached out to genOway to help us in finding the best approach for this project. Their experts had very good understanding of our scientific objective and their thorough analysis helped us to design an effective genetic strategy based on Rosa26 Quick Knockin. The versatility of their conditional expression system helped us to rapidly obtain the proof of concept confirmation before going into more refined tissue-specific expression."

    Stephan Kemp

    Associate Professor at Academic Medical Center (AMC)
 

We have been routinely producing models using Rosa26 and Hprt for over 10 years, which has enabled us to use this approach to create sophisticated models for challenging scientific projects, such as, for example:

  • Humanization for:
    • Human diseases studies
    • Biologics/compound validation
  • Driving disease onset through kinase-dead/active mutants
  • Inducing diseases at certain stages of the animal life cycle
  • Testing protective effects by overexpressing proteins
  • Rescuing Knockout phenotypes
  • Tagging proteins to monitor trafficking and cell fate
  • Single or dual reporter expression for cell tracking

More information on the technical aspects of this approach.

 

A selection of published case studies using our Rosa26 and Hprt models.

Application Scientific Strategy Case study
Mimicking rare human disease Brain-specific expression of a mutated human enzyme Adrenoleukodystrophy mouse model
(PLoS One. 2016) PubMed
Biomarker identification
Drug treatment validation Increase of endogenous enzyme level Attenuation of obesity
(Diabetes. 2015) PubMed
Target validation Expression of self-activated human enzyme Memory deficits in Alzheimer's
(Cell Death Differ. 2014) PubMed
Expression of mutated human protein Acute Myeloid Leukemia mouse model
(Blood. 2013) PubMed
Mimicking inherited disease onset Expression of mutated disease-related protein Lafora disease mouse model
(EMBO Mol Med. 2012) PubMed
Study in vitro vs in vivo effects Block cell membrane protein binding Macrophage chemotaxis
(J Mol Med. 2014) PubMed
Protein trafficking Reporter gene expression under endogenous promoter Antiandrogenic compounds
(PLoS One. 2013) PubMed
Compound characterization
Target characterization Ectopic gene expression Depression mouse model
(PLoS One. 2013) PubMed
Study drug mechanism Antidepressant agents
(Nat Med. 2013) PubMed

 

 

Custom Rosa26 / Hprt Knockin Mouse Services at genOway

genOway provides a one-stop solution for the design, creation and analysis of your custom Knockin mouse model.

Step 1: Scientific evaluation

genOway has a dedicated team of scientists and bioinformatics experts who are able to perform thorough analyses of your project feasibility including:

  • Defining the model's objectives
  • Determining the characteristics of the target gene/locus
  • Analyzing data culled from the literature

Step 2: Design of genetically modified model

We evaluate the possible gene targeting options based on the results of the above scientific evaluation and present the design to the principal investigator with a clear assessment of model features, risks and timelines.

  • Our process guides you toward the best choice for your model design.
  • We provide free and non-binding access to this expertise.

Step 3: Production

genOway has an extensive background in genetically modified mouse technology and will provide you with the highest quality product possible. Four mouse Knockin production options are available. All options include the same dedicated customer support.

Option 1:  Full service

Your project is managed from initial conception to final realization by our in-house transgenesis experts. This level of service includes:

  • Model design, starting from a gene accession number
  • Model creation up to validated heterozygous/homozygous animals
  • Cohort production
  • Phenotyping  (i.e. physiological validation of the model along with a comprehensive array of tests)
Option 2:  Standard service

genOway manages your project from initial conception (starting from a gene accession number) to the delivery of heterozygous animals.

Option 3:  Partial production service

Exit the model creating process at any step of your choosing (such as vector construction or recombined ES cell line creation).

Option 4:  Animal production service

A) By homologous recombination: genOway manages your model creation project, starting from existing targeting vectors or ES cell clones. These vectors or clones are either developed in your lab or available via the International KO Mouse Consortium (IKMC – EUCOMM, KOMP, NorCOMM).

Milestones Full Standard Partial production Animal production
Project evaluation Bioinformatics analysis, bibliography search,
project feasibility assessment
Analysis of the selected strategy,
risk assessment
Model design and targeting
vector construction
x x x  
Recombined ES cells x x optional optional
Blastocyst injection x x optional x
Heterozygous animals x x   x
Homozygous animals x optional   optional
Cohort production x optional   optional
Phenotyping x optional   optional

 

B) By CRISPR/Cas9 nuclease system: genOway manages your model creation project, starting from existing material either developed in your lab or available via third-party providers.

Milestones Full Standard Partial production Animal production
Project evaluation Bioinformatics analysis, bibliography search,
project feasibility assessment
Analysis of the selected strategy,
risk assessment
Model design and nuclease
production
x x x  
Injection into
fertilized embryos
x x optional x
3 independent founders x x   x
Homozygous animals x optional   optional
Cohort production x optional   optional
Phenotyping x optional   optional

Genetic Background

A) For homologous recombination. ES cell lines are currently available for the following genetic backgrounds:
C57BL/6N, C57BL/6J, BALB/c, 129Sv, 129Ola

Our ES cell lines have been validated for germline transmission after 2 rounds of electroporation.

B) For CRISPR/Cas9 nuclease system. We have extensive experience applying CRISPR/Cas9 nuclease technology to virtually any genetic background using specifically adapted protocols and procedures.

Guarantee

Our guarantee comprises:

  • Generation of chimeras
  • ES cell germline transmission
  • Off-target measurement
  • Developmental deadlines
  • Confidentiality
  • Exclusivity of intellectual property
  • Breeding under VAF Elite/SOPF conditions: the health status is issued by professional breeders
  • Line importation and worldwide delivery of the line using a professional transport network

Benefits of Entrusting Your Model Development Project to genOway

Expertise

  • More than 1,500 models generated so far
  • Strong consulting activity provided for model design
  • Thorough project assessment prior to project start

Quality controlled production

  • Robust and validated technologies
  • Clear cut guarantees
  • No waiting list

Project management and reporting

  • A dedicated PhD project manager
  • Rigorous and regular reporting
  • On-site meetings

Unique IP positioning

  • Access to exclusive licenses for key technologies
  • Customer retains all the IP on the model
  • No claim, no royalty, no follow-on rights by genOway