NKI GEMM-ESC archive

The Netherlands Cancer Institute (NKI) has created a unique archive of embryonic stem cells (ESCs) derived from validated genetically engineered mouse models (GEMM) of cancer. These GEMM-ESCs, with often multiple modified alleles, form the basis for further genetic engineering either by Flp-recombinase mediated integration, gene targeting or Crispr/Cas9 to allow for the evaluation of altered target gene expression in a spontaneous tumor model.

The culture and manipulation of GEMM-ESC clones is performed entirely under feeder- and serum-free conditions using the defined N2B27 medium with LIF and the two inhibitors (2i), CHIR99021 and PD0325901, as originally described by the group of Austin Smith, Cambridge, UK. Once the desired genetic alteration is introduced in a GEMM-ESC clone, chimeric mice can be produced using the classic ESC injection protocols that are routinely applied in transgenic facilities. Interestingly, the chimeric mice can be directly used to monitor tumor development as they contain the same genetic alterations as the original GEMM including the altered target gene expression. This GEMM-ESC approach allows for rapid and scalable target gene validation in vivo.

Currently the GEMM-ESC archive contains seven models; four models breast cancer, one for small cell lung cancer (SCLC), one for non-small cell lung cancer (NSCLC) and a model of malignant mesothelioma. A detailed protocol can be found at:

Reference:
Nature Protocols 2015 Oct 15. Using the GEMM-ESC strategy to study gene function in mouse models. Huijbers IJ, Del Bravo J, Bin Ali R, Pritchard C, Braumuller TM, van Miltenburg MH, Henneman L, Michalak EM, Berns A & Jonkers J.

The NKI GEMM-ESC archive is housed at the Mouse Clinic for Cancer and Aging (MCCA) in Amsterdam, the Netherlands. GEMM-ESCs clones are sent under a Material Transfer Agreement (MTA). Shipping costs are for the receiving party and a 500 EUR handling fee is charged per clone.

Validated clones ready for ordering

Validated GEMM-ESC clones / online order form
Direct contact for scientific enquiries and clone distribution is:

Ivo Huijbers PhD
Head of MCCA Transgenic Facility
The Netherlands Cancer Institute
Plesmanlaan 121
1066 CX Amsterdam
The Netherlands

+31 (0)20 512 6172
i.huijbers@nki.nl
www.mccanet.nl

References

Methodology and concept:

Cell Rep. 2016 Jul 19. Transcription Factor NFIB Is a Driver of Small Cell Lung Cancer Progression in Mice and Marks Metastatic Disease in Patients. Semenova EA, Kwon MC, 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.

Genes Dev. 2016 Jun 15. Modeling invasive lobular breast carcinoma by CRISPR/Cas9-mediated somatic genome editing of the mammary gland. Annunziato S, Kas SM, Nethe M, YĆ¼cel 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.

Nature Protocols 2015 Oct 15. Using the GEMM-ESC strategy to study gene function in mouse models. Huijbers IJ, Del Bravo J, Bin Ali R, Pritchard C, Braumuller TM, van Miltenburg MH, Henneman L, Michalak EM, Berns A & Jonkers J.

PNAS. 2015 Jun 22. Selective resistance to the PARP inhibitor olaparib in a mouse model for BRCA1-deficient metaplastic breast cancer. Henneman L, van Miltenburg MH, Michalak EM, Braumuller TM, Jaspers JE, Drenth AP, Grimmerink R, Gogola E, Szuhai K, Schlicker A, Bin Ali R, Pritchard C, Huijbers IJ, Berns A, Rottenberg S, Jonkers J.

EMBO Mol Med. 2014 Feb;6(2):212-25 Rapid target gene validation in complex cancer mouse models using re-derived embryonic stem cells. 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.

Curr Opin Genet Dev. 2012 Feb;22(1):21-7. doi: 10.1016/j.gde.2012.01.004. Epub 2012 Feb 8. Using genetically engineered mouse models to validate candidate cancer genes and test new therapeutic approaches. van Miltenburg MH, Jonkers J.

Bioessays. 2011 Sep;33(9):701-10. doi: 10.1002/bies.201100018. Epub 2011 Jul 7. Rapid validation of cancer genes in chimeras derived from established genetically engineered mouse models. Huijbers IJ, Krimpenfort P, Berns A, Jonkers J.



GEMM development:

Cancer Cell. 2003 Sep;4(3):181-9. Induction of small cell lung cancer by somatic inactivation of both Trp53 and Rb1 in a conditional mouse model. Meuwissen R, Linn SC, Linnoila RI, Zevenhoven J, Mooi WJ, Berns A.

See clone id: TMA10114, TMA10132, TMA10353, TMA11101.


Cancer Cell. 2004 Apr;5(4):375-87. Endogenous oncogenic K-ras(G12D) stimulates proliferation and widespread neoplastic and developmental defects. Tuveson DA, Shaw AT, Willis NA, Silver DP, Jackson EL, Chang S, Mercer KL, Grochow R, Hock H, Crowley D, Hingorani SR, Zaks T, King C, Jacobetz MA, Wang L, Bronson RT, Orkin SH, DePinho RA, Jacks T.

See clone id: TMA11473, IH12059, TMA10819, IH12060, TMA10873

Cancer Cell. 2008 Mar;13(3):261-71. A conditional mouse model for malignant mesothelioma. Jongsma J, van Montfort E, Vooijs M, Zevenhoven J, Krimpenfort P, van der Valk M, van de Vijver M, Berns A.See clone id: TMA10135, TMA10106, IH12076, TMA10668


Proc Natl Acad Sci U S A. 2007 Jul 17;104(29):12111-6. Somatic loss of BRCA1 and p53 in mice induces mammary tumors with features of human BRCA1-mutated basal-like breast cancer. Liu X, Holstege H, van der Gulden H, Treur-Mulder M, Zevenhoven J, Velds A, Kerkhoven RM, van Vliet MH, Wessels LF, Peterse JL, Berns A, Jonkers J.

See clone id: JJ7575, JJ7334, JJ8160

Cancer Cell. 2006 Nov;10(5):437-49. Somatic inactivation of E-cadherin and p53 in mice leads to metastatic lobular mammary carcinoma through induction of anoikis resistance and angiogenesis. Derksen PW, Liu X, Saridin F, van der Gulden H, Zevenhoven J, Evers B, van Beijnum JR, Griffioen AW, Vink J, Krimpenfort P, Peterse JL, Cardiff RD, Berns A, Jonkers J.

See clone id: JJ9965, IH20096, JJ8059, JJ9507, JJ7373, JJ8641, IHM20047

Non-validated clones - nominations for validation are welcome

Non-validated GEMM-ESC clones
Besides the validated GEMM-ESC clones, the complete collection holds an additional 70 clones with different genotypes, reflecting 31 unique alleles. The performance of these clones, i.e. to provide good chimeras with germline transmission, has not been determined, yet. You can nominate a particular GEMM-ESC clone for validation. They can then be prioritized. If you have scientific enquiries or want to nominate a clone for validation, please get in touch with:

Ivo Huijbers PhD
Head of MCCA Transgenic Facility
The Netherlands Cancer Institute
Plesmanlaan 121
1066 CX Amsterdam
The Netherlands

+31 (0)20 512 6172
i.huijbers@nki.nl
www.mccanet.nl

Strain IDAllelesBackgroundTarget tissue
1532_NKIAct-LSL-KrasG12VT/+FVB/NLung
2804_NKI Bap1F/FB6Single
1847_NKI Brca1F/delAG;P53F/FFVB/N;129 
1796_NKI Brca1F/FB6JSingle
1698_NKI Brca1F/F;P53F/FFVB/N  
1915_NKIBrca1F/F;P53F/FB6J 
2218_NKIBrca1F/F;P53F/FFVB/N Breast
1697_NKIBrca1F/F;P53F/F;Act-EGFPT/TFVB/N  
1815_NKIBrca1F/F;P53F/F;Bcrp1-/-FVB/N  
1732_NKIBrca1F/F;P53F/F;Mdr1a/b-/-FVB/N  
1875_NKIBrca1F/InsC;P53F/FFVB/N;129 
1845_NKI Brca1F/MOP1;P53F/F FVB/N;129 
1520_NKI Brca2F/FB6JSingle
294_NKI Brca2F/F;P53F/FFVB/N 
1524_NKI Cdkn2aF/FB6JSingle
1296_NKI Chd9F/FMixSingle
1526_NKI DccF/FB6JSingle
1870_NKI Cdh1F/FFVB/NSingle
2005_NKI Cdh1F/F;P53F/FFVB/N 
296_NKI Cdh1F/F;P53F/F FVB/N 
1622_NKICdh1F/F;P53F/F;RAG1-/-FVB/N  
2181_NKI Cdh1F/F;PtenF/FFVB/N  
1868_NKI Cdh1F/F;T201T/T;SB51T/TFVB/N  
1869_NKI Cdh1F/F;T202T/T;SB51T/TFVB/N  
1847Cre_NKIK14-CreT/+;Brca1F/delAG;P53F/FFVB/N;129Breast
1698Cre_NKI K14-CreT/+;Brca1F/F;P53F/FFVB/N Breast 
1697Cre_NKI K14-CreT/+;Brca1F/F;P53F/F;Act-EGFPT/T FVB/N Breast 
1815Cre_NKI K14-CreT/+;Brca1F/F;P53F/F;Bcrp1-/- FVB/N Breast 
1732Cre_NKI K14-CreT/+;Brca1F/F;P53F/F;Mdr1a/b-/-FVB/N Breast 
1875Cre_NKI K14-CreT/+;Brca1F/InsC;;P53F/FFVB/N;129 Breast 
1845Cre_NKI K14-CreT/+;Brca1F/MOP1;P53F/FFVB/N;129 Breast 
294Cre_NKI K14-CreT/+;Brca2F/F;P53F/FFVB/N Breast 
296Cre_NKI K14-CreT/+;Cdh1F/F;P53F/FFVB/N Breast 
1622Cre_NKI K14-CreT/+;Cdh1F/F;P53F/F;RAG1-/-FVB/N Breast 
1954Cre_NKI K14-CreT/+;P53F/FFVB/N Breast 
1669_NKIKrasLSL-G12D/+B6JLung
2135_NKI KrasLSL-G12D/+P53F/FB6JLung
1683_NKI NF2F/F;Cdkn2aF/FMix(129/FVB)Meso-
thelium
900_NKI Nf2F/F;P16*/*;P53F/FMix(129/FVB) Meso-
thelium
1408_NKI Nf2F/F;P53F/FMix(129/FVB) Meso-
thelium
2973_NKI P15/16BArfF/F;PtenF/FFVB/NLung
2973mTmG_NKI P15/16BArfF/F;PtenF/F;Rosa26-mTmGT/+FVB/N Lung
1800_NKI P53F/FFVB/N Single
1954_NKI P53F/F FVB/N Single
1955_NKI P53F/F FVB/N Single
 2135_NKIP53F/F B6JSingle
1533_NKIPtenF/FB6JSingle
2182_NKI PtenF/F FVB/NSingle
2597_NKI PtenF/F B6JSingle
2597Braf_NKI PtenF/F;BrafCA/+B6J 
1803_NKI Rb1F/FFVB/NSingle
349_NKI Rb1F/F;P53F/F Mix(129/FVB)Lung
2550_NKI Rb1F/F;P53F/F;Col1a1-frtT/+Mix(129/FVB) Lung
2762Col1a_NKI Rb1F/F;P53F/F;Col1a1-frtT/+ Mix(129/FVB) Lung
2762Col1aConf_NKIRb1F/F;P53F/F;Col1a1-frtT/+;Rosa26-LSL-ConfettiT/+ Mix(129/FVB) Lung
2762Conf_NKIRb1F/F;P53F/F;Rosa26-LSL-ConfettiT/+ Mix(129/FVB) Lung
2317_NKIRosa26-CreERT2T/+B6JSingle
2316_NKIRosa26-LSL-LacZT/+B6JSingle
1582_NKIT201T/T;SB51T/TFVB/N 
1583_NKIT202T/T;SB51T/TFVB/N 
 2597Cre_NKITyr-CreERT2T/+;PtenF/F B6JSkin
2597CreBraf_NKITyr-CreERT2T/+;PtenF/F;BrafCA/+B6JSkin
2218Cre_NKIWap-Crehigh/+;Brca1F/F;P53F/FFVB/NBreast
1870Cre_NKIWap-Crehigh/+;Cdh1F/FFVB/NBreast 
2005Cre_NKIWap-Crehigh/+;Cdh1F/F;P53F/FFVB/NBreast
2181Cre_NKIWap-Crehigh/+;Cdh1F/F;PtenF/FFVB/NBreast
1868Cre_NKIWap-Crehigh/+;Cdh1F/F;T201T/T;SB51T/TFVB/NBreast
1869Cre_NKIWap-Crehigh/+;Cdh1F/F;T202T/T;SB51T/TFVB/NBreast
1955Cre_NKIWap-Crehigh/+;P53F/FFVB/NBreast
2182Cre_NKIWap-Crehigh/+;PtenF/FFVB/NBreast
Alleles
AlleleGeneModificationPubMed ID
Act-LSL-KrasG12VKirsten RasActin promoter Lox-STOP-lox G12V 11641780
Bap1Bap1floxedunpublished
Bcrp1Abcg2knockout12429862
BrafProto-oncogene B-RafKnockin Lox-STOP-lox-V600E mutation17299132
Brca1Breast Cancer 1, early onsetfloxed17626182
Brca2Breast Cancer 2, early onset floxed11694875
Cdh1Cadherin 1floxed17097565
Cdn2aCyclin Dependent Kinase Inhibitor p16 and p19floxed11544530
Chd9Chromodomain helicase DNA binding protein 9 (Obx)floxedunpublished
DccDeleted in Colorectal Carcinomafloxed22358843
Del AGBrca1185 delAG mutationunpublished
EGFPEnhanced GFPActin promoter9867352
InscBrca15382InsC mutationunpublished
K14-CreCre recombinaseCytokeratin 14 promoter1694875
KrasLSL-G12DKirsten RasKnockin Lox-STOP-lox-G12D mutation15093544
Mdr1a/bAbcb1a and Abcb1bKnockout9108099
MOPBrca1exon 5 mutant22172724
mTmGtomato red and GFPRosa26 promoter; floxed tomato red17868096
Nf2Neurofibromatosis type 2floxed10887156
P15/16BArfp15, p16 and p19p15 and p16 knockout; p19Arf floxed17713536
P16P16Pointmutant p16 (p16 function loss)11544530
P53Tumor protein p53floxed11694875
PtenPhosphatase and tensin homology (Pten)floxed12091320
RAG1Recombination Activating Gene 1knockout1547488
Rb1Retinoblastoma proteinfloxed7925271
Rosa26-CreERT2CreERT2Rosa26 ubiquitous promoter17456738
Rosa26-LSL-Confetti Lox-STOP-Lox Confetti reporter (Brainbow2.1) Rosa26 ubiquitous promoter20887898
Rosa26-LSL-LacZLox-STOP-Lox b-galactosidaseRosa26 ubiquitous promoter9916792
SB51Sleeping beauty T2/Onc transposon system 16015333
T201T2/onc line 68 16015333
Wap-CreCre recombinaseWhey Acidic Protein promoter (High)21282721