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Conditional mouse lung cancer models using adenoviral or lentiviral delivery of Cre recombinase

Nature Protocols volume 4, pages 1064–1072 (2009)Cite this article

Abstract

The development of animal models of lung cancer is critical to our understanding and treatment of the human disease. Conditional mouse models provide new opportunities for testing novel chemopreventatives, therapeutics and screening methods that are not possible with cultured cell lines or xenograft models. This protocol describes how to initiate tumors in two conditional genetic models of human non-small cell lung cancer (NSCLC) using the activation of oncogenic K-ras alone or in combination with the loss of function of p53. We discuss methods for sporadic expression of Cre in the lungs through engineered adenovirus or lentivirus, and provide a detailed protocol for the administration of the virus by intranasal inhalation or intratracheal intubation. The protocol requires 1–5 min per mouse with an additional 30–45 min to set-up and allow for the recovery of mice from anesthesia. Mice may be analyzed for tumor formation and progression starting 2–3 weeks after infection.

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Figure 1: Intratracheal infection technique.
Figure 2: Intranasal infection technique.
Figure 3: Survival is reduced in the K-rasLSL-G12D/+;p53fl/fl (KP) model compared with K-rasLSL-G12D/+ (K) model.
Figure 4: Tumor progression and histopathological phenotype in K-rasLSL-G12D/+;p53fl/fl (KP) model.

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Acknowledgements

We would like to thank Carla Kim and Amber Woolfenden for originally training the authors to carry out the intratracheal intubation technique. The technique was implemented in our lab with help from Kwok-Kin Wong and Samanthi Perera. We would like to thank Trudy Oliver and Peter Sandy for providing data to compile the Kaplan–Meier survival curves as well as Denise Crowley and Roderick Bronson for providing key histological and pathological advice. We also thank Keara Lane, Etienne Meylan, Eric Snyder and Anne Deconinck for reviewing this protocol. This work was supported by funding from the Howard Hughes Medical Institute, the NCI (including a Cancer Center Support grant) and the Ludwig Center for Molecular Oncology at MIT. T.J. is the David H. Koch Professor of Biology and a Daniel K. Ludwig Scholar. Research was conducted in compliance with the Animal Welfare Act Regulations and other Federal statutes relating to animals and experiments involving animals and adheres to the principles set forth in the Guide for Care and Use of Laboratory Animals, National Research Council, 1996.

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Author notes

  1. Michel DuPage and Alison L Dooley: These authors contributed equally to this work.

Authors and Affiliations

  1. Koch Institute for Integrative Cancer Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Michel DuPage, Alison L Dooley & Tyler Jacks

  2. Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Tyler Jacks

Authors
  1. Michel DuPage
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  2. Alison L Dooley
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  3. Tyler Jacks
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Contributions

M.D., A.L.D. and T.J. wrote the manuscript.

Corresponding author

Correspondence to Tyler Jacks.

Supplementary information

Supplementary Fig. 1: Genetically controlled events in mouse model of lung cancer.

Engineering of LoxP elements and a stop element allow for the controlled expression of oncogenic K-ras and the loss of p53 function after Cre expression. (PDF 224 kb)

Supplementary Fig. 2: Preparation of the Exel Safelet IV catheter for intratracheal infection.

Upon opening the Exel Safelet IV catheter, the needle is exposed (a). Slide the catheter over the end of the needle to completely cover the tip (b) and the Exel Safelet IV catheter is now ready to use. (PDF 7475 kb)

Supplementary Fig. 3: Recovery following intranasal or intratracheal infection.

Mice can be placed under a heat lamp (a) or on a glove filled with warm water (b) to recover following anesthesia in the biosafety hood. CAUTION All experiments should be done in accordance with protocols approved by the Institutional Animal Care and Use Committee. (PDF 5898 kb)

Supplementary Video 1: Intratracheal intubation technique.

CAUTION All experiments should be done in accordance with protocols approved by the Institutional Animal Care and Use Committee. (MOV 12504 kb)

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DuPage, M., Dooley, A. & Jacks, T. Conditional mouse lung cancer models using adenoviral or lentiviral delivery of Cre recombinase. Nat Protoc 4, 1064–1072 (2009). https://doi.org/10.1038/nprot.2009.95

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