Name: novel method of plasmid dna delivery to mouse bladder urothelium by electroporation
Uploaded: 2018-05-03T14:30:00.000+00:00
Duration: 6 min 38 s
Description: Watch this Scientific Journal Video about Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation at JoVE.com

We thank Professor Bin Chen, Dr. Yue Zhang, and Kendy Hoang for help with setting up the electroporation system. This work was supported by grants from the Santa Cruz Cancer Benefit Group and NIH to Z.A.W.

All procedures described here were performed in accordance with the guidelines and regulations from the Institutional Animal Care and Use Committee (IACUC) of University of California Santa Cruz.
1. Plasmid and Tools Preparation
<ol>
	<li>For each bladder to transfect, prepare at least 20 &#181;L of DNA plasmid (1 &#181;g/&#181;L).</li>
	<li>Add 1 &#181;L of Trypan Blue to the 20 &#181;L of plasmid solution in a 1.5 mL microcentrifuge tube. Pipette to mix well.</li>
	<li>Autoclave all surgical instruments and sterilize the workspace with 70% ethanol. Spray surgical instruments and gloves with 70% ethanol frequently or use a glass bead sterilizer&#160;when performing the following steps to maintain sterile conditions.</li>
</ol>
2. Anaesthetizing Animal
<ol>
	<li>Use a table top anesthesia system to anesthetize the animal with isoflurane. Turn on the O2 tank and adjust the oxygen flowmeter to 1 L/min.</li>
	<li>Place an adult female C57BL/6J mouse in the induction chamber. Turn on and adjust the isoflurane vaporizer to 3% for induction. The mouse should be in anesthesia within 2 min.&#160;Administer buprenorphine analgesic (0.1 mg/kg) subcutaneously upon removal of the mouse from the induction chamber for preemptive analgesia. 
	NOTE: In this protocol, female mice are used, as they are easier to work with during urethra catheterization. The protocol also works for male mice, but extra caution is needed when performing step 3.</li>
	<li>Apply ophthalmic ointment to the eyes of the animal to prevent dryness. Place the anaesthetized mouse facing up on a heating pad with its nose in the nose cone. Turn the air circuit switch to let isoflurane go through the nose cone.</li>
	<li>Restrain the head and the nose cone with adhesive tape. Adjust the isoflurane vaporizer to 2% for maintenance.</li>
	<li>Restrain the limbs with adhesive tape. Perform toe-pinch tests to make sure the animal is in deep anesthesia and then shave the abdominal area.</li>
</ol>
3. Urine Depletion and Bladder Rinsing
<ol>
	<li>Apply lubricant to the catheter (24G, length of 2.11 cm, outer diameter of 0.045 cm) and ensure that its outer surface is fully covered.</li>
	<li>Insert the catheter into the urethra opening and slowly push it until it reaches the bladder, at which time the urine should flow out through the catheter. Gently press the abdomen to help urine depletion.
	<ol>
		<li>Check if the catheter is inserted correctly into the bladder by observing automatic urine out-flow. Avoid piercing the urethra and bladder wall, and apply lubricant multiple times if needed.</li>
	</ol>
	</li>
	<li>Discard the urine with a pipette into a waste beaker.</li>
	<li>Pipette 80 &#181;L of phosphate buffered saline (PBS) into the outer end of the catheter, with the other end still in the bladder. Carefully attach a 1 mL syringe to the catheter. Gently push the syringe to inject the PBS into the bladder. Leave some PBS in the catheter to avoid creating air bubbles in the bladder.</li>
	<li>Remove the syringe. Wait for PBS to drain out of the bladder. Gently press the abdomen to evacuate PBS.</li>
	<li>Discard the PBS in the catheter with a pipette into the waste beaker.</li>
	<li>Repeat PBS washing (steps 3.4 - 3.6) for two more times. 
	NOTE: It is essential to work gently during these washing steps. Blood in the catheter or failure of automatic out-flow of liquid usually indicates the urethra is pierced through. It is also important to avoid introducing air bubbles, as they will decrease the electroporation efficiency.</li>
</ol>
4. Plasmid Delivery
<ol>
	<li>Apply 70% ethanol to the mouse abdomen and use a scalpel to make a vertical incision of 1 cm to open the abdominal skin above the bladder.</li>
	<li>Pipette at least 20 &#181;L of plasmid plus Trypan Blue solution into the outer end of the catheter, and attach the syringe.</li>
	<li>Inject the plasmid solution into the bladder. If the bladder turns blue, the injection is successful. Leave some liquid in the catheter to avoid creating air bubbles in the bladder.</li>
	<li>Grab the external urethral orifice using tweezers, and then remove the catheter and syringe. Use a string to tighten the external urethral orifice. Make two loops with the string and then tie with two knots. 
	NOTE: Tightening of the urethral is important for preventing backflow of plasmid liquid.</li>
	<li>Turn on the electroporation generator with the following parameters: 33V, 50 ms working time, and 950 ms interval time.</li>
	<li>Hold the bladder with tweezers and pinch the bladder with two electrodes. Push the foot pedal to perform the electroporation. 
	NOTE: The electric pulses are set to occur 5 times with short intervals after each pedal push. The mouse may twitch in this process. Pushing the foot pedal more than once can increase the delivery efficiency, but too many electric pulses can damage the tissue integrity of the urothelium.
	<ol>
		<li>Avoid any contact between the tweezers and the electrodes, as this will generate a spark.</li>
	</ol>
	</li>
</ol>
5. Recovery
<ol>
	<li>Suture the abdominal and skin opening with sterile surgical suture and clips. Apply iodine onto the wound.</li>
	<li>Remove the animal from the isoflurane system. Administer buprenorphine analgesic (0.1 mg/kg) subcutaneously to alleviate post-surgical pain.</li>
	<li>Keep the animal on the heating pad and return it to the home cage after full recovery. Check the animal at least once daily for normal mobility, drinking and feeding behaviors and no signs of infection until the incision is healed and then remove the clips. Administer subsequent injections of buprenorphine analgesic if the animal displays pain symptom such as reduced grooming, increased aggressiveness, and vocalization.</li>
</ol>

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The authors declare no competing interest.

Electroporation enhances the cell membrane permeability and is a powerful technology for gene electrotransfer16. Gene delivery into living rodents by electroporation has been frequently used in neurobiology fields17,18,19,20,21. Its potential applications in many other organs have not been extensively explored. To our knowledge, our met...

Genetically engineered mouse models (GEMMs) have been playing an essential role in expanding our knowledge about animal development, gene functions in vivo, and mechanisms of disease progression1,2. Germline GEMMs are traditionally developed in two ways. The first is the creation of transgenic mice by pronuclear injection of DNA plasmid followed by the transplantation of zygotes into pseudopregnant females. The other is the generation of knock-out/knock-in mice by homologous recombination in embryonic stem cells and the development of chimeras. Conditional knockout of genes in specific tissue or cell type of interest involves the breeding of genetically engineered alleles with Cre and flox sites for multiple generations. The whole process can be expensive and time-consuming, especially if the goal is to target multiple genes tissue-specifically. Recently, gene-editing technologies such as clustered regularly interspaced short palindromic repeats (CRISPR) have been applied to several organs of the mice to induce tissue-specific genetic alterations for autochthonous cancer modeling3,4,5,6,7 or correct disease mutations in situ8,9. In these studies, the delivery of the gRNA vectors was usually achieved through adenoviral or lentiviral infection of the organ or hydrodynamic tail-vein injection. The relative ease of delivery of the CRISPR components to the lung and liver has greatly improved the convenience and efficiency of cancer modeling in these organs compared to the traditional Cre-Lox based mouse models.
The bladder urothelium is where most bladder tumors originate. The delivery of DNA vectors to the bladder urothelium for cancer modeling or gene therapy is hampered by a glycosaminoglycan layer on the apical urothelial surface, which acts as a barrier for adherence of infectious viruses10,11. Several pretreatment agents such as Syn3 and dodecyl-&#946;-d-maltoside have been used to disrupt this barrier and enhance adenovirus infection efficiency12,13,14. Whether adeno-associated viruses (AAVs) can effectively infect the bladder has not been reported. Overall, a non-viral based delivery method would be desirable. Here, we describe a novel method developed in the lab for efficient DNA plasmid delivery to the mouse urothelium through urethra catheterization and electroporation. Because our approach does not involve chemical pretreatment of the glycosaminoglycan layer or virus production, it significantly simplifies the delivery of DNA plasmids into the mouse bladder urothelium, and should facilitate various in vivo studies of bladder diseases.

<table><tbody><tr><td>BD 1mL TB Syringe</td><td>Fisher</td><td>148232E</td><td></td></tr><tr><td>Buprenorphine</td><td>Sigma</td><td>B9275-50MG</td><td></td></tr><tr><td>Dumont Tweezers</td><td>Roboz Surgical Instr</td><td>RS-5005</td><td>Treat with 70% ethanol before surgical use</td></tr><tr><td>ECM830 SQ Wave Electroporator</td><td>Harvard Apparatus</td><td>450052</td><td></td></tr><tr><td>Exel International Disposable Safelet I.V. Catheters</td><td>Fisher Scientific</td><td>14-841-21</td><td>24G, 2.11 cm length, 0.045 cm outer diameter</td></tr><tr><td>Extra Fine Micro Dissecting Scissors</td><td>Roboz Surgical Instr</td><td>RS-5135</td><td>Treat with 70% ethanol before surgical use</td></tr><tr><td>Isoflurane</td><td>Vet one</td><td>501017</td><td></td></tr><tr><td>K&amp;amp;H Heated Resting Mat for Small Animals, 9 By 12 Inches</td><td>Amazon</td><td>n/a</td><td>Cover the heat mat with paper towels</td></tr><tr><td>Ophthalmic ointment</td><td>Fisher</td><td>NC0849514</td><td></td></tr><tr><td>PBS, pH7.4</td><td>Life Technologies</td><td>10010049</td><td></td></tr><tr><td>Pipet tips 200 &amp;micro;L</td><td>USA Scientific</td><td>1111-0206</td><td></td></tr><tr><td>Pipet Tips, Universal Fit; 0.1 to 10 &amp;micro;L</td><td>Fisher</td><td>02707438(CS)</td><td></td></tr><tr><td>PIPETMAN NEO P2</td><td>Gilson</td><td>F144561</td><td></td></tr><tr><td>PIPETMAN NEO P200N</td><td>Gilson</td><td>F144565</td><td></td></tr><tr><td>plasmid CAG-GFP</td><td>Addgene</td><td>16664</td><td></td></tr><tr><td>Platinum tweezertrode 5 mm</td><td>Harvard Apparatus</td><td>450489</td><td>5 mm diameter</td></tr><tr><td>Scissors</td><td>Roboz Surgical Instr</td><td>RS-5880</td><td>Treat with 70% ethanol before surgical use</td></tr><tr><td>String cord</td><td>Amazon</td><td>n/a</td><td>Mandala Crafts 150D 210D 0.8mm 1mm leather sewing stitching flat waxed thread string cord</td></tr><tr><td>Tape, Scotch</td><td>Fisher</td><td>19047257</td><td></td></tr><tr><td>Therio-gel Veterinary Lubricant</td><td>PBS animal health</td><td>353-736</td><td></td></tr><tr><td>Trypan Blue Stain</td><td>Life Technologies</td><td>15250061</td><td></td></tr><tr><td>V-1 Table top system anesthesia</td><td>VetEquip</td><td>901806</td><td></td></tr><tr><td>Vicryl violet suture 18&quot; FS-2 cutting 5-0</td><td>Fisher</td><td>NC0578475</td><td></td></tr><tr><td>Walgreens Povidone Iodine 10%</td><td>Walgreens</td><td>953982</td><td></td></tr><tr><td>Wound clip</td><td>Fisher</td><td>018045</td><td></td></tr><tr><td>Wound clip applier</td><td>Fisher</td><td>01804</td><td></td></tr></tbody></table>

novel method of plasmid dna delivery to mouse bladder urothelium by electroporation

Genetically engineered mouse models (GEMMs) are extremely valuable in revealing novel biological insights into the initiation and progression mechanisms of human diseases such as cancer. Transgenic and conditional knockout mice have been frequently used for gene overexpression or ablation in specific tissues or cell types in vivo. However, generating germline mouse models can be time-consuming and costly. Recent advancements in gene editing technologies and the feasibility of delivering DNA plasmids by viral infection have enabled rapid generation of non-germline autochthonous mouse cancer models for several organs. The bladder is an organ that has been difficult for viral vectors to access, due to the presence of a glycosaminoglycan layer covering the urothelium. Here, we describe a novel method developed in lab for efficient delivery of DNA plasmids into the mouse bladder urothelium in vivo. Through intravesical instillation of pCAG-GFP DNA plasmid and electroporation of surgically exposed bladder, we show that the DNA plasmid can be delivered specifically into the bladder urothelial cells for transient expression. Our method provides a fast and convenient way for overexpression and knockdown of genes in the mouse bladder, and can be applied to building GEMMs of bladder cancer and other urological diseases.

To demonstrate the success of gene delivery into the bladder urothelium, we used the pCAG-GFP15 plasmid for electroporation. 20 &#181;L of plasmid and Trypan Blue solution was injected through the urethra and 5 electric pulses were administered. After 48 h, we dissected the mouse bladder and observed patches of GFP fluorescence under a dissection fluorescence microscope, whereas a negative control bladder that did not undergo electroporation showed no GFP signal (<...

Watch this Scientific Journal Video about Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation at JoVE.com

Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation

We describe a novel method for the delivery of DNA plasmid into the urothelial cells of mouse bladder in vivo through urethra catheterization and electroporation. It offers a fast and convenient way for generating autochthonous mouse models of bladder diseases.

Genetically engineered mouse models (GEMMs) are extremely valuable in revealing novel biological insights into the initiation and progression mechanisms ...

novel-method-plasmid-dna-delivery-to-mouse-bladder-urothelium

Research

JoVE Journal

Biology

8.4K Views.  University of California, Santa Cruz. We describe a novel method for the delivery of DNA plasmid into the urothelial cells of mouse bladder in vivo through urethra catheterization and electroporation. It offers a fast and convenient way for generating autochthonous mouse models of bladder diseases.

Video: Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation

Mouse Bladder Wall Injection

Mouse bladder wall injection is a useful technique to orthotopically study bladder phenomena, including stem cell, smooth muscle, and cancer biology. Before starting injections, the surgical area must be cleaned with soap and water and antiseptic solution. Surgical equipment must be sterilized before use and between each animal. Each mouse is placed under inhaled isoflurane anesthesia (2-5% for induction, 1-3% for maintenance) and its bladder exposed by making a midline abdominal incision with scissors. If the bladder is full, it is partially decompressed by gentle squeezing between two fingers. The cell suspension of interest is intramurally injected into the wall of the bladder dome using a 29 or 30 gauge needle and 1 cc or smaller syringe. The wound is then closed using wound clips and the mouse allowed to recover on a warming pad. Bladder wall injection is a delicate microsurgical technique that can be mastered with practice.

Mouse bladder wall injection is a useful approach to orthotopically study bladder stem cell and cancer biology. This delicate microsurgical method can be mastered with careful technique and practice.

Mouse bladder wall injection is a useful technique to orthotopically study bladder phenomena, including stem cell, smooth muscle, and cancer biology. ...

Medicine

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

In order to understand the function of genes expressed in specific region of the developing brain, including signaling molecules and axon guidance molecules, local gene transfer or knock- out is required. Gene targeting knock-in or knock-out into local regions is possible to perform with combination with a specific CRE line, which is laborious, costly, and time consuming. Therefore, a simple transfection method, an in utero electroporation technique, which can be performed with short time, will be handy to test the possible function of candidate genes prior to the generation of transgenic animals 1,2. In addition to this, in utero electroporation targets areas of the brain where no specific CRE line exists, and will limit embryonic lethality 3,4. Here, we present a method of in utero electroporation combining two different types of electrodes for simple and convenient gene transfer into target areas of the developing brain. First, a unique holding method of embryos using an optic fiber optic light cable will make small embryos (from E9.5) visible for targeted DNA solution injection into ventricles and needle type electrodes insertion to the targeted brain area 5,6. The patterning of the brain such as cortical area occur at early embryonic stage, therefore, these early electroporation from E9.5 make a big contribution to understand entire area patterning event. Second, the precise shape of a capillary prevents uterine damage by making holes by insertion of the capillary. Furthermore, the precise shape of the needle electrodes are created with tungsten and platinum wire and sharpened using sand paper and insulated with nail polish 7, a method which is described in great detail in this protocol. This unique technique allows transfection of plasmid DNA into restricted areas of the brain and will enable small embryos to be electroporated. This will help to, open a new window for many scientists who are working on cell differentiation, cell migration, axon guidance in very early embryonic stage. Moreover, this technique will allow scientists to transfect plasmid DNA into deep parts of the developing brain such as thalamus and hypothalamus, where not many region-specific CRE lines exist for gain of function (GOF) or loss of function (LOF) analyses.

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

A gene transfer method into the developing mouse brain is described by using a unique surgical method and special shape of electrodes. This unique technique allows transfection of plasmid DNA temporally and spatially, which will aid many neuroscientists in studying brain development.

In order to understand the function of genes expressed in specific region of the developing brain, including signaling molecules and axon guidance ...

Neuroscience

An Orthotopic Bladder Cancer Model for Gene Delivery Studies

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are needed. The tumor microenvironment can significantly influence tumor development and therapy response. It is therefore often desirable to grow tumor cells in the organ from which they originated. This protocol describes an orthotopic model of bladder cancer, in which MB49 murine bladder carcinoma cells are instilled into the bladder via catheterization. Successful tumor cell implantation in this model requires disruption of the protective glycosaminoglycan layer, which can be accomplished by physical or chemical means. In our protocol the bladder is treated with trypsin prior to cell instillation. Catheterization of the bladder can also be used to deliver therapeutics once the tumors are established. This protocol describes the delivery of an adenoviral construct that expresses a luciferase reporter gene. While our protocol has been optimized for short-term studies and focuses on gene delivery, the methodology of mouse bladder catheterization has broad applications.

Implantation of cancer cells into the organ of origin can serve as a useful preclinical model to evaluate novel therapies. MB49 bladder carcinoma cells can be grown within the bladder following intravesical instillation. This protocol demonstrates catheterization of the mouse bladder for the purpose of tumor implantation and adenoviral delivery.

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are ...

In Vivo Microinjection and Electroporation of Mouse Testis

This video and article contribution gives a comprehensive description of microinjection and electroporation of mouse testis in vivo. This particular transfection technique for testicular mouse cells allows the study of unique processes in spermatogenesis.
The following protocol focuses on transfection of testicular mouse cells with plasmid constructs. Specifically, we used the reporter vector pEGFP-C1, which expresses enhanced green fluorescent protein (eGFP) and also the pDsRed2-N1 vector expressing red fluorescent protein (DsRed2). Both encoded reporter genes were under the control of the human cytomegalovirus immediate-early promoter (CMV).
For performing gene transfer into mouse testes, the reporter plasmid constructs are injected into testes of living mice. To that end, the testis of an anaesthetized animal is exposed and the site of microinjection is prepared. Our preferred place of injection is the efferent duct, with the ultimately connected rete testis as the anatomical transport route of the spermatozoa between the testis and the epididymis. In this way, the filling of the seminiferous tubules after microinjection is excellently managed and controlled due to the use of stained DNA solutions. After observing a sufficient filling of the testis by its colored tubule structure, the organ is electroporated. This enables the transfer of the DNA solution into the testicular&#160;cells. Following 3 days of incubation, the testis is removed and investigated under the microscope for green or red&#160;fluorescence, illustrating transfection success.
Generally, this protocol can be employed for delivering DNA- or RNA- constructs into living mouse testis in order to (over)express or knock down genes, facilitating in vivo gene function analysis. Furthermore, it is suitable for studying reporter constructs or putative gene regulatory elements. Thus, the main advantages of the electroporation technique are fast performance in combination with low effort as well as the moderate technical equipment and skills required compared to alternative techniques.

In Vivo Microinjection and Electroporation of Mouse Testis

This article describes microinjection and electroporation of mouse testis in&#160;vivo as a&#160;transfection technique for testicular mouse cells to study unique processes of spermatogenesis. The presented protocol involves steps of glass capillary preparation, microinjection via the efferent duct, and transfection by electroporation.

This video and article contribution gives a comprehensive description of microinjection and electroporation of mouse testis in vivo. This particular ...

Electroporation of Plasmid DNA into Mouse Skeletal Muscle

Transient gene expression modulation in murine skeletal muscle by plasmid electroporation is a useful tool for assessing normal and pathological physiology. Overexpression or knockdown of target genes enables investigators to manipulate individual molecular events and, thus, better understand the mechanisms that impact muscle mass, muscle metabolism, and contractility. In addition, electroporation of DNA plasmids that encode fluorescent tags allows investigators to measure changes in subcellular localization of proteins in skeletal muscle in vivo. A key functional assessment of skeletal muscle includes the measurement of muscle contractility. In this protocol, we demonstrate that whole muscle contractility studies are still possible after plasmid DNA injection, electroporation, and gene expression modulation. The goal of this instructional procedure is to demonstrate the step-by-step method of DNA plasmid electroporation into mouse skeletal muscle to facilitate uptake and expression in the myofibers of the tibialis anterior and extensor digitorum longus muscles, as well as to demonstrate that skeletal muscle contractility is not compromised by injection and electroporation.

Electroporation of plasmid DNA into skeletal muscle is a viable method to modulate gene expression without compromising muscle contractility in mice.

Transient gene expression modulation in murine skeletal muscle by plasmid electroporation is a useful tool for assessing normal and pathological ...

Expression of Transgenes in Native Bladder Urothelium Using Adenovirus-Mediated Transduction

In addition to forming a high-resistance barrier, the urothelium lining the renal pelvis, ureters, bladder, and proximal urethra is hypothesized to sense and transmit information about its environment to the underlying tissues, promoting voiding function and behavior. Disruption of the urothelial barrier, or its sensory/transducer function, can lead to disease. Studying these complex events is hampered by lack of simple strategies to alter gene and protein expression in the urothelium. Methods are described here that allow investigators to generate large amounts of high-titer adenovirus, which can then be used to transduce rodent urothelium with high efficiency, and in a relatively straightforward manner. Both cDNAs and small interfering RNAs can be expressed using adenoviral transduction, and the impact of transgene expression on urothelial function can be assessed 12 h to several days later. These methods have broad applicability to studies of normal and abnormal urothelial biology using mouse or rat animal models.

Methods are described for the generation of large amounts of recombinant adenoviruses, which can then be used to transduce the native rodent urothelium allowing for expression of transgenes or downregulation of endogenous gene products.

In addition to forming a high-resistance barrier, the urothelium lining the renal pelvis, ureters, bladder, and proximal urethra is hypothesized to sense ...

Electroporation-Mediated In Vitro Gene Delivery: An Electric Pulse Technique to Introduce Fluorescent Reporter Protein-Encoding Plasmids Into Cultured Cells

Source: Lo, J. C. W. et al., <a href="https://www.jove.com/v/63283">Efficient and Cost Effective Electroporation Method to Study Primary Cilium-Dependent Signaling Pathways in the Granule Cell Precursor.</a>&#160;J. Vis. Exp.&#160; (2021)
This video demonstrates the electroporation technique that introduces fluorescent protein-encoding plasmid DNA into mouse primary cerebellar granule cell precursors.

Dostarczanie genów in vitro za pośrednictwem elektroporacji: technika impulsów elektrycznych do wprowadzania fluorescencyjnych plazmidów kodujących białko reporterowe do hodowanych komórek

Source: Lo, J. C. W. et al., Efficient and Cost Effective Electroporation Method to Study Primary Cilium-Dependent Signaling Pathways in the Granule Cell ...

JoVE Encyclopedia of Experiments

Biological Techniques

Gene Transfer Techniques

Transfection

Urine Depletion and Bladder Rinsing in Mouse Model: A Catheter-based Procedure to Empty and Wash Murine Urinary Bladder

Source: Yu, C. et al. <a href="https://www.jove.com/video/57649" target="_blank">Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation.</a> J. Vis. Exp. (2018)
In this video, we describe a catheter-based procedure to empty and rinse the urinary bladder of a mouse model. The prepped mouse can then be used for further experimental procedures.

Wyczerpanie moczu i płukanie pęcherza moczowego w modelu mysim: procedura oparta na cewniku do opróżniania i mycia mysiego pęcherza moczowego

Source: Yu, C. et al. Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation. J. Vis. Exp. (2018)
In this video, we describe ...

Cancer Research

Urinary Tract Cancer

In vivo studies

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

Source: Chuan, Y. et al. <a href="https://www.jove.com/video/57649" target="_blank">Novel method of plasmid DNA delivery to mouse bladder urothelium by electroporation.</a> J. Vis. Exp.&#160; (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.

Dostarczanie plazmidów DNA do nabłonka dróg moczowych pęcherza myszy: procedura oparta na elektroporacji mająca na celu skuteczne dostarczanie plazmidów DNA do komórek urotelialnych pęcherza myszy w celu wygenerowania modeli modyfikowanych genetycznie

Source: Chuan, Y. et al. Novel method of plasmid DNA delivery to mouse bladder urothelium by electroporation. J. Vis. Exp.&#160; (2018)
This video ...

Hydrodynamic Renal Pelvis Injection: A Technique to Directly Deliver DNA into Kidney Cells of Murine Model

Source: Woodard, L. E. et al. <a href="https://www.jove.com/video/56324" target="_blank">Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney.</a> J. Vis. Exp. (2018)
In this video, we present the protocol for hydrodynamic injection of plasmids into the renal pelvis of the mouse kidney for nucleic acid transfection. This technique allows organ-specific delivery of plasmids by utilizing the principle of high hydrostatic pressure, which forcefully drives the plasmids into the specific site without causing tissue injury.

Hydrodynamiczne wstrzyknięcie do miednicy nerkowej: technika bezpośredniego dostarczania DNA do komórek nerkowych modelu mysiego

Source: Woodard, L. E. et al. Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney. J. Vis. Exp. (2018)
In this video, ...

Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation

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In Vivo Microinjection and Electroporation of Mouse Testis

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Expression of Transgenes in Native Bladder Urothelium Using Adenovirus-Mediated Transduction

Dostarczanie genów in vitro za pośrednictwem elektroporacji: technika impulsów elektrycznych do wprowadzania fluorescencyjnych plazmidów kodujących białko reporterowe do hodowanych komórek

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Hydrodynamiczne wstrzyknięcie do miednicy nerkowej: technika bezpośredniego dostarczania DNA do komórek nerkowych modelu mysiego