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
Paul Krimpenfort PhD
The Netherlands Cancer Institute
Plesmanlaan 121
1066 CX Amsterdam
The Netherlands
+31 (0)20 512 1284
p.krimpenfort@nki.nl
www.mccanet.nl
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, 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.
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
Paul Krimpenfort
The Netherlands Cancer Institute
Plesmanlaan 121
1066 CX Amsterdam
The Netherlands
+31 (0)20 512 1284
p.krimpenfort@nki.nl
www.mccanet.nl
| Strain ID | Alleles | Background | Target tissue |
| 1532_NKI | Act-LSL-KrasG12VT/+ | FVB/N | Lung |
| 2804_NKI | Bap1F/F | B6 | Single |
| 1847_NKI | Brca1F/delAG;P53F/F | FVB/N;129 | |
| 1796_NKI | Brca1F/F | B6J | Single |
| 1698_NKI | Brca1F/F;P53F/F | FVB/N | |
| 1915_NKI | Brca1F/F;P53F/F | B6J | |
| 2218_NKI | Brca1F/F;P53F/F | FVB/N | Breast |
| 1697_NKI | Brca1F/F;P53F/F;Act-EGFPT/T | FVB/N | |
| 1815_NKI | Brca1F/F;P53F/F;Bcrp1-/- | FVB/N | |
| 1732_NKI | Brca1F/F;P53F/F;Mdr1a/b-/- | FVB/N | |
| 1875_NKI | Brca1F/InsC;P53F/F | FVB/N;129 | |
| 1845_NKI | Brca1F/MOP1;P53F/F | FVB/N;129 | |
| 1520_NKI | Brca2F/F | B6J | Single |
| 294_NKI | Brca2F/F;P53F/F | FVB/N | |
| 1524_NKI | Cdkn2aF/F | B6J | Single |
| 1296_NKI | Chd9F/F | Mix | Single |
| 1526_NKI | DccF/F | B6J | Single |
| 1870_NKI | Cdh1F/F | FVB/N | Single |
| 2005_NKI | Cdh1F/F;P53F/F | FVB/N | |
| 296_NKI | Cdh1F/F;P53F/F | FVB/N | |
| 1622_NKI | Cdh1F/F;P53F/F;RAG1-/- | FVB/N | |
| 2181_NKI | Cdh1F/F;PtenF/F | FVB/N | |
| 1868_NKI | Cdh1F/F;T201T/T;SB51T/T | FVB/N | |
| 1869_NKI | Cdh1F/F;T202T/T;SB51T/T | FVB/N | |
| 1847Cre_NKI | K14-CreT/+;Brca1F/delAG;P53F/F | FVB/N;129 | Breast |
| 1698Cre_NKI | K14-CreT/+;Brca1F/F;P53F/F | FVB/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/F | FVB/N;129 | Breast |
| 1845Cre_NKI | K14-CreT/+;Brca1F/MOP1;P53F/F | FVB/N;129 | Breast |
| 294Cre_NKI | K14-CreT/+;Brca2F/F;P53F/F | FVB/N | Breast |
| 296Cre_NKI | K14-CreT/+;Cdh1F/F;P53F/F | FVB/N | Breast |
| 1622Cre_NKI | K14-CreT/+;Cdh1F/F;P53F/F;RAG1-/- | FVB/N | Breast |
| 1954Cre_NKI | K14-CreT/+;P53F/F | FVB/N | Breast |
| 1669_NKI | KrasLSL-G12D/+ | B6J | Lung |
| 2135_NKI | KrasLSL-G12D/+P53F/F | B6J | Lung |
| 1683_NKI | NF2F/F;Cdkn2aF/F | Mix(129/FVB) | Meso- thelium |
| 900_NKI | Nf2F/F;P16*/*;P53F/F | Mix(129/FVB) | Meso- thelium |
| 1408_NKI | Nf2F/F;P53F/F | Mix(129/FVB) | Meso- thelium |
| 2973_NKI | P15/16BArfF/F;PtenF/F | FVB/N | Lung |
| 2973mTmG_NKI | P15/16BArfF/F;PtenF/F;Rosa26-mTmGT/+ | FVB/N | Lung |
| 1800_NKI | P53F/F | FVB/N | Single |
| 1954_NKI | P53F/F | FVB/N | Single |
| 1955_NKI | P53F/F | FVB/N | Single |
| 2135_NKI | P53F/F | B6J | Single |
| 1533_NKI | PtenF/F | B6J | Single |
| 2182_NKI | PtenF/F | FVB/N | Single |
| 2597_NKI | PtenF/F | B6J | Single |
| 2597Braf_NKI | PtenF/F;BrafCA/+ | B6J | |
| 1803_NKI | Rb1F/F | FVB/N | Single |
| 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_NKI | Rb1F/F;P53F/F;Col1a1-frtT/+;Rosa26-LSL-ConfettiT/+ | Mix(129/FVB) | Lung |
| 2762Conf_NKI | Rb1F/F;P53F/F;Rosa26-LSL-ConfettiT/+ | Mix(129/FVB) | Lung |
| 2317_NKI | Rosa26-CreERT2T/+ | B6J | Single |
| 2316_NKI | Rosa26-LSL-LacZT/+ | B6J | Single |
| 1582_NKI | T201T/T;SB51T/T | FVB/N | |
| 1583_NKI | T202T/T;SB51T/T | FVB/N | |
| 2597Cre_NKI | Tyr-CreERT2T/+;PtenF/F | B6J | Skin |
| 2597CreBraf_NKI | Tyr-CreERT2T/+;PtenF/F;BrafCA/+ | B6J | Skin |
| 2218Cre_NKI | Wap-Crehigh/+;Brca1F/F;P53F/F | FVB/N | Breast |
| 1870Cre_NKI | Wap-Crehigh/+;Cdh1F/F | FVB/N | Breast |
| 2005Cre_NKI | Wap-Crehigh/+;Cdh1F/F;P53F/F | FVB/N | Breast |
| 2181Cre_NKI | Wap-Crehigh/+;Cdh1F/F;PtenF/F | FVB/N | Breast |
| 1868Cre_NKI | Wap-Crehigh/+;Cdh1F/F;T201T/T;SB51T/T | FVB/N | Breast |
| 1869Cre_NKI | Wap-Crehigh/+;Cdh1F/F;T202T/T;SB51T/T | FVB/N | Breast |
| 1955Cre_NKI | Wap-Crehigh/+;P53F/F | FVB/N | Breast |
| 2182Cre_NKI | Wap-Crehigh/+;PtenF/F | FVB/N | Breast |
| Allele | Gene | Modification | PubMed ID |
| Act-LSL-KrasG12V | Kirsten Ras | Actin promoter Lox-STOP-lox G12V | 11641780 |
| Bap1 | Bap1 | floxed | unpublished |
| Bcrp1 | Abcg2 | knockout | 12429862 |
| Braf | Proto-oncogene B-Raf | Knockin Lox-STOP-lox-V600E mutation | 17299132 |
| Brca1 | Breast Cancer 1, early onset | floxed | 17626182 |
| Brca2 | Breast Cancer 2, early onset | floxed | 11694875 |
| Cdh1 | Cadherin 1 | floxed | 17097565 |
| Cdn2a | Cyclin Dependent Kinase Inhibitor p16 and p19 | floxed | 11544530 |
| Chd9 | Chromodomain helicase DNA binding protein 9 (Obx) | floxed | unpublished |
| Dcc | Deleted in Colorectal Carcinoma | floxed | 22358843 |
| Del AG | Brca1 | 185 delAG mutation | unpublished |
| EGFP | Enhanced GFP | Actin promoter | 9867352 |
| Insc | Brca1 | 5382InsC mutation | unpublished |
| K14-Cre | Cre recombinase | Cytokeratin 14 promoter | 1694875 |
| KrasLSL-G12D | Kirsten Ras | Knockin Lox-STOP-lox-G12D mutation | 15093544 |
| Mdr1a/b | Abcb1a and Abcb1b | Knockout | 9108099 |
| MOP | Brca1 | exon 5 mutant | 22172724 |
| mTmG | tomato red and GFP | Rosa26 promoter; floxed tomato red | 17868096 |
| Nf2 | Neurofibromatosis type 2 | floxed | 10887156 |
| P15/16BArf | p15, p16 and p19 | p15 and p16 knockout; p19Arf floxed | 17713536 |
| P16 | P16 | Pointmutant p16 (p16 function loss) | 11544530 |
| P53 | Tumor protein p53 | floxed | 11694875 |
| Pten | Phosphatase and tensin homology (Pten) | floxed | 12091320 |
| RAG1 | Recombination Activating Gene 1 | knockout | 1547488 |
| Rb1 | Retinoblastoma protein | floxed | 7925271 |
| Rosa26-CreERT2 | CreERT2 | Rosa26 ubiquitous promoter | 17456738 |
| Rosa26-LSL-Confetti | Lox-STOP-Lox Confetti reporter (Brainbow2.1) | Rosa26 ubiquitous promoter | 20887898 |
| Rosa26-LSL-LacZ | Lox-STOP-Lox b-galactosidase | Rosa26 ubiquitous promoter | 9916792 |
| SB51 | Sleeping beauty T2/Onc transposon system | 16015333 | |
| T201 | T2/onc line 68 | 16015333 | |
| Wap-Cre | Cre recombinase | Whey Acidic Protein promoter (High) | 21282721 |