Delivery of DNA Plasmids into Urothelium of Mouse Bladder: An Electroporation-based Procedure to Efficiently Deliver DNA Plasmids into Urothelial Cells of Murine Bladder to Generate Genetically Engineered Models

Published: April 30, 2023

Abstract

Source: Chuan, Y. et al. Novel method of plasmid DNA delivery to mouse bladder urothelium by electroporation. J. Vis. Exp.  (2018)

This video demonstrates the delivery of DNA plasmid into the urothelium of the mouse bladder using a combination of urethral catheterization and electroporation techniques. The described procedure allows the generation of mouse bladder cancer models.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Plasmid and Tools Preparation For each bladder to transfect, prepare at least 20 µL of DNA plasmid (1 µg/µL). Add 1 µL of Trypan Blue to the 20 µL of plasmid solution in a 1.5 mL microcentrifuge tube. Pipette to mix well. Autoclave all surgical instruments and sterilize the w…

Disclosures

The authors have nothing to disclose.

Materials

BD 1mL TB Syringe Fisher 148232E
Buprenorphine Sigma B9275-50MG
Dumont Tweezers Roboz Surgical Instr RS-5005 Treat with 70% ethanol before surgical use
ECM830 SQ Wave Electroporator Harvard Apparatus 450052
Exel International Disposable Safelet I.V. Catheters Fisher Scientific 14-841-21 24G, 2.11 cm length, 0.045 cm outer diameter
Extra Fine Micro Dissecting Scissors Roboz Surgical Instr RS-5135 Treat with 70% ethanol before surgical use
Isoflurane Vet one 501017
K&H Heated Resting Mat for Small Animals, 9 by 12 Inches Amazon Cover the heat mat with paper towels
Ophthalmic ointment Fisher NC0849514
PBS, pH 7.4 Life Technologies 10010049
Pipet tips 200 µL USA Scientific 1111-0206
Pipet Tips, Universal Fit; 0.1 to 10 µL Fisher 02707438(CS)
PIPETMAN NEO P2 Gilson F144561
PIPETMAN NEO P200N Gilson F144565
plasmid CAG-GFP Addgene 16664
Platinum tweezertrode 5 mm Harvard Apparatus 450489 5 mm diameter
Scissors Roboz Surgical Instr RS-5880 Treat with 70% ethanol before surgical use
String cord Amazon Mandala Crafts 150D 210D 0.8mm 1mm leather sewing stitching flat waxed thread string cord
Tape, Scotch Fisher 19047257
Therio-gel Veterinary Lubricant PBS animal health 353-736
Trypan Blue Stain Life Technologies 15250061
V-1 Table top system anesthesia VetEquip 901806
Vicryl violet suture 18" FS-2 cutting 5-0 Fisher NC0578475
Walgreens Povidone Iodine 10% Walgreens 953982
Wound clip Fisher 18045
Wound clip applier Fisher 1804

Tags

Play Video

Cite This Article
Delivery of DNA Plasmids into Urothelium of Mouse Bladder: An Electroporation-based Procedure to Efficiently Deliver DNA Plasmids into Urothelial Cells of Murine Bladder to Generate Genetically Engineered Models. J. Vis. Exp. (Pending Publication), e20564, doi: (2023).

View Video