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

Published: April 30, 2023

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 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 canc…

Representative Results

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 GFP reporter allele in this mouse. (Apctm2Rak, Krastm4Tyj, Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze<…

Açıklamalar

The authors have nothing to disclose.

Materials

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 GmbH 188271/227270
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 DICKINSON 304000
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 Vaporizer Parkland Scientific  V3000PS/PK
Cellulose swabs  Lohmann & Rauscher Deutschland  13356
Insulin syringe EMG 1 mL (with 30G cannula) B. Braun Melsungen AG 9161627S
Fine Bore Tubing (bore: 0.28 mm/ diameter: 0.61mm) Smiths Medical Deutschland 8 800/100/100
Micro-Adson Forceps Fine Science Tools 11018-12
Iris Scissor – ToughCut Fine Science Tools 14058-11
Olsen-Hegar Needle Holder Fine Science Tools 12002-12
AutoClip Kit Fine Science Tools 12020-00
PDS Z1012H 6/0 C1 (surgical suture) Johnson & Johnson Medical GmbH Z1012H
Curved Micro Serrefine Vascular Clamp Fine Science Tools 18055-05
Fogarty Spring Clips Edwards CDSAFE 6
Hot Plate 062 Labotect  13854
Isis – Hair shaver Aesculap – Braun
Cold Light Fountain XENON 175 Karl Storz 20132101-1 Karl Storz Coloview System Mainz
Fiber Optic Light Cable Karl Storz 69495NL Karl Storz Coloview System Mainz
TRICAM Three-Chip Camera Head Karl Storz 20221030 Karl Storz Coloview System Mainz
TRICAM SLII Camera Control Unit Karl Storz 20223011-1 Karl Storz Coloview System Mainz
15" Flat Screen Monitor EndoVue Karl Storz 9415NN Karl Storz Coloview System Mainz
HOPKINS Straight Forward Telescope diameter 1.9 mm; length 10 cm autoclavable fiber optic light transmission incorporated Karl Storz 64301AA
Protection and Examination Sheath Karl Storz 61029C

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Bu Makaleden Alıntı Yapın
Segmental Adeno-Cre Infection: A Technique to Generate Isolated Colorectal Cancer using Genetically Engineered Mouse Models. J. Vis. Exp. (Pending Publication), e20338, doi: (2023).

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