GEMreport

An Advanced Academic Reporting System For Genetically Engineered Mouse

Welcome to MCCA Transgenic Facility

The Mouse Clinic for Cancer and Aging (MCCA) is part of the Netherlands Cancer Institute (NKI) and provides all services related to the generation of genetically engineered mice using both our in-house developed GEMM-ESC pipeline and the CRISPR-Cas9 genome editing technology.

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To consult or request services, we kindly ask you to ceate an account. You can then fill in the consultation form and provide some basic information on the type of genetically engineered mouse model you would like to develop. Upon submission, you will be contacted shortly.

SERVICES WE PROVIDE

The MCCA Transgenic Facility provides the typical services related to the generation and cryopreservation of mutant mouse strains supplemented with several unique services. A list of individual services is shown below organized by type of approach.

GEMM-ESC pipeline – UNIQUE

 Aim: delivery of high-quality chimeras

CRISPR-Cas9 gene editing in zygotes

  • Construct design
  • Cloning of gRNA constructs
  • ssODN design for homology directed repair
  • Zygote micro-injection
  • Screening of founder mice (F0)
  • Screening of F1 generation
 Aim: delivery of sequence verified F1 mice

Classic gene targeting

  • Construct design
  • Cloning of targeting construct
  • Development of screening PCR
  • ESC targeting
  • Screening by PCR and Southern blot
  • Blastocyst micro-injections
 Aim: delivery of high-quality chimeras

Cryopreservation

  • Sperm freezing via EMMA protocol
  • Embryo freezing (only for NKI researchers)

WHO WE ARE

The MCCA Transgenic Facility is part of the NKI, located in Amsterdam, and has a long history in the generation of genetically modified mouse models.

Ambition

The NKI is a comprehensive cancer center consisting of a research institute and a specialized clinic. One of the pillars in its research is disease modeling in mice. The NKI has clustered the expertise in mouse model generation and preclinical testing in one core facility, named the Mouse Clinic for Cancer and Aging research, MCCA in short. Within the MCCA there are two facilities. The MCCA Transgenic Facility generates genetically engineered mouse strains and archives these mouse lines by cryogenic storage of embryos or sperm. The Mouse Cancer Clinic performs preclinical intervention studies. The MCCA’s ambition is to have a long-term impact on scientific research in cancer and aging and to contribute to the validation of new treatments that improve patient survival and quality of live.

State-of-the-art infrastructure

The infrastructure of the MCCA Transgenic facility is elaborate and state-of-the-art. The activities are spread over 5 dedicated labs:

  • A transgenic lab
  • Two large mouse holding rooms
  • A molecular biology lab
  • An ESC culture lab
  • An RNA production lab

The transgenic lab has four micro-injection stations for injecting GEMM-ESCs or CRISPR in early embryos. All mice are kept in individually ventilated cages under strict hygienic standards.

Recent publications

  • Huijbers IJ. Generating Genetically Modified Mice: A Decision Guide. Methods in molecular biology. 2017:1642:1-19.
  • Pritchard CEJ, Kroese LJ, Huijbers IJ. Direct Generation of Conditional Alleles Using CRISPR/Cas9 in Mouse Zygotes.Methods in molecular biology. 2017:1642:21-35.
  • Ferone G, Song JY, Sutherland KD, Bhaskaran R, Monkhorst K, Lambooij JP, Proost N, Gargiulo G, Berns A. SOX2 Is the Determining Oncogenic Switch in Promoting Lung Squamous Cell Carcinoma from Different Cells of Origin. Cancer Cell. 2016:30:519-532.
  • 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. 2016;16:631-43.
  • 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. Modeling invasive lobular breast carcinoma by CRISPR/Cas9-mediated somatic genome editing of the mammary gland. Genes & Development. 2016;30:1470-80.
  • Pilzecker B, Buoninfante OA, Pritchard C, Blomberg OS, Huijbers IJ, van den Berk P, Jacobs H. PrimPol prevents APOBEC/AID family mediated DNA mutagenesis. Nucleic Acids Research. 2016;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. Using the GEMM-ESC strategy to study gene function in mouse models. Nature Protocols. 2015;10:1755-85.
  • 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. Selective resistance to the PARP inhibitor olaparib in a mouse model for BRCA1-deficient metaplastic breast cancer. PNAS 2015;112:8409-14.

FULL-SERVICE MODEL

The MCCA Transgenic Facility generates genetically engineered mice for academic researchers with full-service covering the design and all hands-on steps as DNA engineering, ESC manipulation and micro-injection.

Full-service

The procedure starts with an initial brainstorm to determine the optimal mouse model. Next a practical approach is devised to generate this mouse model with the highest chance of success. Once agreed by all parties, the MCCA Transgenic Facility will perform all the individual development steps to generate the requested genetically modified mouse. The first- or second-generation mice, including a method for screening, are delivered to the customers who will do the breeding, validations and perform the actual experiments. The customer is advised to archive the newly validated mouse model by freezing of sperm.


full service

Conditions

The MCCA Transgenic Facility provides fee-for-service under not-for-profit conditions. The mice are generated in a Specific Pathogen Free (SPF) environment. The MCCA has a best effort obligation and can not guarantee delivery of the requested mouse line due unforeseen complications.

MCCA TRANSGENIC FACILITY


Manager

Ivo Huijbers PhD

i.huijbers@nki.nl
P: +31 (0)20 512 6172

Location

Plesmanlaan 121
1066 CX Amsterdam
The Netherlands