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Method Article

Segmental Adeno-Cre Infection: A Technique to Generate Isolated Colorectal Cancer using Genetically Engineered Mouse Models

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April 30th, 2023

In This Article

Abstract

Source: Betzler, A. M., et al. A Genetically Engineered Mouse Model of Sporadic Colorectal Cancer. J. Vis. Exp. (2017).

This video describes the generation of a colorectal mouse model by segmental adeno-cre infection. The infected colon segment leads to tumorigenesis within this segment of the mucosa, resulting in adenomas ultimately progressing to invasive and metastatic carcinoma. This model is an attractive platform for studies of cancer biology and preclinical therapeutic trials.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Local Tumor Induction via Surgical Adeno-cre Infection

  1. Preparation of animals for surgery
    NOTE: Virtually any conditional ("floxed") mutation can be induced via the here described method. The use of mutations in genes relevant to colorectal cancer, such as Apc, Kras, or Tp53, is recommended. The efficiency of Cre recombination is dependent on the size of the construct to be excised. Large floxed sequences are ....

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Results

Endoscopic views of the esophagus, showing tissue variations and potential abnormalities.
Figure 1. Colonoscopic Images of Colorectal Tumors (Conditional Alleles of the Given Animal in Brackets).
A. Normal distal colon. B. Early adenoma 2 weeks after adeno-cre infection. Note the green color due to a .......

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Disclosures

No conflicts of interest declared.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
Dulbecco's Phosphate Buffered Saline Life Technologies GmbH 14190169
Trypsin-EDTA (0.25%, PhenolRed) Life Technologies GmbH 25200072
Normal saline 0.9% (E154) Serumwerk Bernburg AG 10013
Aqua ad injectabilia B. Braun Melsungen AG 235144
Ad5CMV-Cre (adenovirus, c = 2E +11 PFU/mL) Gene Transfer Vector Core University of Iowa
15 mL, 50 mL centrifuge tubes  Greiner Bio-One GmbH188271/227270
Eppendorf tubes 1.5 mL/ 2 mL Sarstedt AG & Co. 72,695,400
Petri dish PS 100/15 mm (sterile, Nuclon) Fisher Scientific GmbH 10508921/ NUNC150350
1 mL Syringe (without dead volume) - Injekt-F SOLO Braun/neoLab 194291661
30G injection needle BECTON DICKINSON304000
Sevoflurane (Sevoflurane AbbVie) AbbVie Germany GmbH & Co. KG
Medical oxygen Air Liquide Medical GmbH
Buprenorphine (Temgesic)Indivior Eu Ltd.
Bepanthen - ophthalmic ointment Bayer Vital GmbH 10047757
Table Top Research Anesthesia Machine x/O2 Flush w/ Sevoflurane VaporizerParkland Scientific V3000PS/PK
Cellulose swabs Lohmann & Rauscher Deutschland 13356
Insulin syringe EMG 1 mL (with 30G cannula)B. Braun Melsungen AG9161627S
Fine Bore Tubing (bore: 0.28 mm/ diameter: 0.61mm)Smiths Medical Deutschland 8800/100/100
Micro-Adson ForcepsFine Science Tools11018-12
Iris Scissor - ToughCutFine Science Tools14058-11
Olsen-Hegar Needle HolderFine Science Tools12002-12
AutoClip KitFine Science Tools12020-00
PDS Z1012H 6/0 C1 (surgical suture)Johnson & Johnson Medical GmbHZ1012H
Curved Micro Serrefine Vascular ClampFine Science Tools18055-05
Fogarty Spring ClipsEdwardsCDSAFE 6
Hot Plate 062Labotect 13854
Isis - Hair shaverAesculap - Braun
Cold Light Fountain XENON 175Karl Storz20132101-1Karl Storz Coloview System Mainz
Fiber Optic Light CableKarl Storz69495NLKarl Storz Coloview System Mainz
TRICAM Three-Chip Camera HeadKarl Storz20221030Karl Storz Coloview System Mainz
TRICAM SLII Camera Control UnitKarl Storz20223011-1Karl Storz Coloview System Mainz
15" Flat Screen Monitor EndoVueKarl Storz9415NNKarl Storz Coloview System Mainz
HOPKINS Straight Forward Telescope diameter 1.9 mm; length 10 cm autoclavable fiber optic light transmission incorporatedKarl Storz64301AA
Protection and Examination SheathKarl Storz61029C

Tags

Colorectal Cancer Mouse ModelGenetically Engineered Mouse ModelCre Recombinase EnzymeLoxP InsertsAdenoviral Suspension InjectionColonoscopy Tumor MonitoringDistal Colon SegmentationTrypsin EDTA TreatmentAbdominal Cavity Exposure