RNA Interference Mice & Rats

With its ability to silence almost any gene, RNA interference (RNAi) is an invaluable tool for identifying and validating potential drug targets in vivo.

RNAi technology is ideal if:
  • Your gene of interest is difficult to knock out
  • You need an inducible or reversible knockdown model
  • You want to simultaneously knockdown two genes
  • You want to mimic the action of an antagonistic drug
At Taconic Biosciences, we’ve helped researchers around the world generate RNAi mouse and rat models to move their studies forward.

Taconic's RNAi models allow for the ubiquitous reversible induction of RNA-mediated gene knockdowns at selected time points. In the absence of RNAi inducer (doxycycline), the tet-repressor protein binds to the tet-operator control element and effectively inhibits the expression of short hairpin RNA (shRNA) from the H1 promoter.

When doxycycline is administered through an animal's feed or drinking water, it binds to the tet-repressor protein and blocks its interaction with the H1 promoter. This is followed by H1 promoter activation, shRNA expression, and effective RNA-mediated gene knockdown. Withdrawal of doxycycline inhibits shRNA expression, resulting in pre-knockdown levels of target gene expression.

Inducible RNAi Rat Models:

Inducible RNAi Rat The absence of suitable rat ES cells has made rat gene targeting studies highly challenging. Through the use of transgenic technology, however, Taconic is pleased to offer its customers inducible RNAi rat models. Pronuclear injection methodology has enabled Taconic scientists to microinject an inducible/reversible shRNA construct into the pronucleus of rat embryos. Following microinjection, embryos are transferred into pseudopregnant female rats. Resultant pups are then screened for the presence of the shRNA expression transgene. Expression of the shRNA in transgenic rats is induced by the administration of doxycycline in food or water. Withdrawal of the inducer enables inhibition of shRNA expression.

Double Gene shRNA Knockdown

  • Study the genetic interaction of two targets by knocking down two genes at the same time.
  • Express two different shRNAs that recognize the same mRNA molecule in genes that are difficult to knockdown with a single shRNA.
  • Target a disease-causing mutation in one allele while leaving the normal allele intact.

  • Double Shrna Knockdown Chain Reporter Fluc Graphs Double Reporter Gene

    qPCR Validation Service

    Taconic provides expression analysis service for all major mouse tissues. The experimental setup consists of a control group and an shRNA targeted group of mice. Both groups are further subdivided into a doxycycline-treated and an untreated group.

    As a standard, Taconic uses a total of 12 mice for one experiment (six control and six shRNA targeted mice).

    Gpcr Validation Graph
    Example of a tissue expression analysis of two RNAi lines and one control line (Stopcontrol). Leakiness in uninduced brain samples may be observed.

    Tissues Routinely Dissected
    Brain (mix) Cerebellum
    Cortex Hippocampus
    Hypothalamus Striatum
    Entire Aorta Heart
    Muscle (Thigh) Epididymal White Adipose (E-WAT)
    Brown Adipose Tissue (BAT) Lung
    Kidney Colon
    Small Intestine Trachea
    Prostate Thyroid
    Bone Marrow Spleen
    Whole Blood Thymus
    Liver Ovary
    Testis Uterus
Give us your target gene and we’ll create your custom RNAi mouse models by:
  1. Identifying a set of suitable shRNAs against your target gene
  2. Cloning a shRNA cassette into an exchange vector and introducing it into the ROSA26 locus of ES cells via recombinase mediated cassette exchange (RMCE)
  3. Examining the ability of the shRNA expression cassette to effectively induce gene knockdown in vitro
  4. If the targeted gene is not expressed in ES cells, introducing an exchange vector containing both the shRNA expression cassette and the target cDNA into ES cells by RMCE
  5. Conducting knockdown analysis in vitro
  6. When effective gene knockdown is achieved in vitro, microinjecting ES cells containing the suitable shRNA expression cassette into mouse blastocysts and then transferring the blastocysts into pseudopregnant female mice
  7. Breeding resultant chimeras to generate mice heterozygous for the knockdown construct

Inducible In Vivo RNAi Gene Knockdown

  • Induce and reverse ubiquitous gene knockdowns at selected time points
  • Achieve a knockdown of more than 70% in most mouse tissues
  • Induce a gene knockdown at any time by administering doxycycline through the animal's feed or drinking water
  • Integrate a single cassette carrying a shRNA with a guaranteed in vitro knockdown of at least 70% into the well-defined ROSA26 locus
  • Breed and type later generations simply and efficiently using just one cassette that contains all selection markers and regulatory elements of the system
  • Wait only 38 weeks to generate models on a pure C57BL/6NTac background

Taconic has generated more than 200 different RNAi mouse models. Your dedicated Taconic project manager will leverage the company's extensive knowledge base to help you choose your mouse model and assist you in planning model validation.

Packages include:

  • Documentation and verification of PCR-based genotype protocol by Taconic Molecular Analysis.
  • Coordinated transfer of donor males to Taconic's breeding facility in Germantown, NY.
  • Confirmation of genotype of donor males by PCR-based genotyping.
  • Rapid Expansion using C57BL/6NTac donor females.
  • Holding of excess donor males until offspring from rapid expansion are weaned.
  • Initial post-derivation comprehenisve health testing (IHMS™-52).
  • PCR based genotyping of derived offspring.
  • Holding of derived pups from weaning until ready for shipment (4 weeks) at the Isolator Breeding Solutions facility.
  • Initial cohort of animals ready for shipment beginning at 5 weeks of age.
    • Large package 18-20/sex/genotype
    • Small package 8-10/sex/genotype
    • Genotypes include heterozygous and wild type

Additional package details:

  • Package ends when initial cohort is ready for shipping. Post package breeding and fees will be based on your breeding plan.
  • Package includes initial health testing only, ongoing health testing can be done for an additional charge.
  • Shipping charges from Taconic to customer are not included in package.
  • Package assumes receipt of 4 donor males from Taconic, additional services will be charged at catalog prices.
  • Pricing assumes genetic modification is not sex linked, Mendelian ratios will be obtained, 80% of female breeders will deliver live pups that survive until weaning, and an average litter size of 5 pups. Additional charges may apply if these conditions are not met.
Gerrits H, Paradé MC, Koonen-Reemst AM, Bakker NE, Timmer-Hellings L, Sollewijn Gelpke MD, Gossen JA. (2014) Reversible infertility in a liver receptor homologue-1 (LRH-1)-knockdown mouse model. Reprod Fertil Dev. 26(2):293-306.

Michalak EM, Nacerddine K, Pietersen A, Beuger V, Pawlitzky I, Cornelissen-Steijger P, Wientjens E, Tanger E, Seibler J, van Lohuizen M, Jonkers J. (2013) Polycomb group gene Ezh2 regulates mammary gland morphogenesis and maintains the luminal progenitor pool. Stem Cells. 31(9):1910-20.

Vink PM, Smout WM, Driessen-Engels LJ, de Bruin AM, Delsing D, Krajnc-Franken MA, Jansen AJ, Rovers EF, van Puijenbroek AA, Kaptein A, Nolte MA, Garritsen A, van Eenennaam H. (2013) In vivo knockdown of TAK1 accelerates bone marrow proliferation/differentiation and induces systemic inflammation. PLoS One. 8(3):e57348.