The Netherlands Cancer Institute has a very strong history in
the generation and validation of mouse models of human cancer. Many
conditional mouse strains for tumor suppressor alleles often lost
or mutated in human cancer, such as Trp53, PTEN and Cdkn2a, have
been generated here and are routinely used worldwide. In that
context, the transgenic core facility accommodates all activities
required to generate genetically modified animals, from DNA and
Embryonic Stem Cell (ESC) services to transgenic services, such as
ESC microinjection, zygote injection as well as cryopreservation.
State-of-the-art technologies, such as ESC re-derivation from
complex models and genome editing, are also offered.
The transgenic core facility is part of the Mouse Clinic for
Cancer and Aging (MCCA). The MCCA is a collaboration of the
Netherlands Cancer Institute and the European Research Institute
for the Biology of Ageing in Groningen, The Netherlands. The MCCA
is funded by a Roadmap grant from the Netherlands Organisation for
Scientific Research (NWO).
Websites: www.mccanet.nl or gem.nki.nl
Generating genetically engineered mice is a complex process that
requires a good strategy and detailed planning. In consultation
with the researcher we provide advice on the strategy and construct
design for generating the mouse strain tailor-made to answer a
biological question. Depending on the complexity and workload a
practical plan is devised to reach that goal and in some case might
involve participation of the researcher in cloning and/or ESC
culture. Our services are listed below and include cryopreservation
of new and existing strains.
- Generation of targeting and Flp-in constructs
- Design screening PCRs for targeting constructs
- Design and cloning of CRISPR gRNA plasmids
- Design ssODN for homology directed repair
- ESC re-derivation and cryogenic storage
- ESC quality testing
- Targeting and Flp-in of ESCs
- PCR and Southern blot screening
- ESC micro-injection/generation of chimeric mice
- Zygote injection; RNA and/or DNA injection
- Cryopreservation - sperm
- Cryopreservation - embryo
Huijbers IJ. Generating Genetically Modified Mice: A
Methods in Molecular Biology. 2017:1642:1-19.
Pritchard CEJ, Kroese LJ, Huijbers IJ. Direct
Generation of Conditional Alleles Using CRISPR/Cas9 in Mouse
Methods in Molecular Biology. 2017:1642:21-35.
Semenova EA, Kwon M-C, Monkhorst K, Song JY, Bhaskaran R, Krijgsman
O, Kuilman T, Peters D, Buikhuisen WA, Smit EF, Pritchard C,
Cozijnsen M, van der Vliet J, Zevenhoven J, Lambooij JP, Proost N,
van Montfort E, Velds A, Huijbers IJ, Berns A. Transcription factor
NFIB is a driver of Small Cell Lung Cancer progression in mice and
marks metastatic disease in patients. Cell Reports.
Annunziato S, Kas SM, Nethe M, Yücel H, Del Bravo J, Pritchard C,
Bin Ali R, van Gerwen B, Siteur B, Drenth AP, Schut E, van de Ven
M, Boelens MC, Klarenbeek S, Huijbers IJ, van Miltenburg MH,
Jonkers J. (2016) Modeling invasive lobular breast carcinoma by
CRISPR/Cas9-mediated somatic genome editing of the mammary gland.
Genes Dev. 30:1470-80.
Pilzecker B, Buoninfante OA, Pritchard C, Blomberg OS, Huijbers
IJ, van den Berk PC, Jacobs H. (2016) PrimPol prevents APOBEC/AID
family mediated DNA mutagenesis. Nucleic Acids Res. 44: 4734-44.
Huijbers IJ, Del Bravo J, Bin Ali R, Pritchard C, Braumuller TM,
van Miltenburg MH, Henneman L, Michalak EM, Berns A & Jonkers
J. (2015) Using the GEMM-ESC strategy to study gene function in
mouse models. Nature Protocols
Henneman L, van Miltenburg MH, Michalak EM, Braumuller TM, Jaspers
JE, Drenth AP, de Korte-Grimmerink R, Gogola E, Szuhai K, Schlicker
A, Bin Ali R, Pritchard C, Huijbers IJ, Berns A, Rottenberg S,
Jonkers J. (2015) Selective resistance to the PARP inhibitor
olaparib in a mouse model for BRCA1-deficient metaplastic breast
2015 Jun 22.
Bin Ali R, van der Ahé F, Braumuller TM, Pritchard C, Krimpenfort
P, Berns A, Huijbers IJ. (2014) Improved pregnancy and birth rates
with routine application of nonsurgical embryo transfer. Transgenic Res.
Huijbers IJ, Bin Ali R, Pritchard C, Cozijnsen M, Kwon MC, Proost
N, Song JY, de Vries H, Badhai J, Sutherland K, Krimpenfort P,
Michalak EM, Jonkers J, Berns A (2014) Rapid target gene validation
in complex cancer mouse models using re-derived embryonic stem
Mol Med 6: 212-25.
Huijbers IJ, Krimpenfort P, Berns A and Jonkers J. (2011) Rapid
validation of cancer genes in chimeras derived from established
genetically engineered mouse models. Bioessays 33, 701-710