エレクトロポレーション システムのセットアップのヘルプ、箱陳教授、博士越張とケンディ ホアンに感謝します。この作品は、Z.A.W. にサンタ ・ クルスがん給付グループと NIH からの補助金によって支えられました。

ガイドラインと規制機関動物ケアおよび使用委員会 (IACUC) カリフォルニア大学サンタクルスからに従ってここで説明したすべての手順を行った。
1 プラスミドとツールの準備
<ol><li>それぞれの膀胱を使って、DNA のプラスミッド (1 μ g/μ L) の少なくとも 20 μ L を準備します。</li>
	<li>1.5 mL 容マイクロ チューブにおけるプラスミド溶液 20 μ L にトリパン ブルーの 1 μ L を追加します。よく混ぜてピペット。</li>
	<li>オートクレーブすべて手術機器および 70% のエタノールでワークスペースを滅菌します。スプレー手術器具や手袋の 70% エタノール頻繁または無菌状態を維持するために次の手順を実行するときにガラス ビーズ滅菌器を使用します。</li>
</ol>2. 麻酔動物
<ol><li>イソフルランと動物を麻酔するのにテーブル トップ麻酔システムを使用します。O2タンクをオンにし、1 L/分の酸素流量計を調整します。</li>
	<li>誘導室に大人の女性 c57bl/6 j マウスを配置します。オンにし、誘導のための 3% にイソフルラン気化器を調整します。マウスは、先制鎮痛法の誘導室からマウスの除去時に皮下麻酔 2 分管理ブプレノルフィン鎮痛剤 (0.1 mg/kg) 以内でなければなりません。 注: このプロトコルでメスのマウスとして使用されます、尿道カテーテル留置中に操作しやすくしています。雄マウスのプロトコルも動作しますが、ステップ 3 を実行するときに余分な注意が必要です。</li>
	<li>乾燥を防ぐためには、動物の目に眼軟膏を適用します。鼻の円錐形の鼻と加熱パッドの上を向いてハイパーカプニアのマウスを置きます。イソフルラン鼻の円錐形を通過させる空気回路のスイッチをオンにします。</li>
	<li>頭と粘着テープで鼻の円錐形を抑制します。メンテナンスのための 2% にイソフルラン気化器を調整します。</li>
	<li>粘着テープで手足を拘束します。動物は深い麻酔で、その後の腹部領域を剃るようにつま先ピンチ テストを実行します。</li>
</ol>3. 尿の枯渇と膀胱洗浄
<ol><li>(24 G、2.11 cm の長さ、0.045 cm の外径) カテーテルに潤滑剤を適用し、その外表面が完全に覆われていることを確認します。</li>
	<li>尿道開口部にカテーテルを挿入し、その時点で尿をカテーテルを通じて流出、膀胱に達するまでゆっくりと差し込みます。尿枯渇のため腹部を優しく押します。<ol>
<li>かどうか、カテーテルが正しく挿入されて、膀胱に自動尿流出を観察することによって確認してください。尿道と膀胱壁の穿孔を回避し、必要な場合は、潤滑剤を複数回適用。</li>
	</ol>
</li>
	<li>廃棄物ビーカーにピペットで尿を破棄します。</li>
	<li>まだ膀胱内のもう一方の端をカテーテルの外側の端にリン酸緩衝生理食塩水 (PBS) の 80 μ L をピペットします。慎重に 1 mL 注射器をカテーテルに接続します。PBS を膀胱内に注入する注射器を軽く押します。カテーテルを膀胱内に空気の泡を作成するを避けるためにいくつかの PBS を残します。</li>
	<li>注射器を削除します。膀胱から排出する PBS を待ちます。PBS を避難する腹部を優しく押します。</li>
	<li>廃棄物のビーカーにピペットでカテーテルで PBS を破棄します。</li>
	<li>さらに 2 回の PBS (ステップ 3.4 3.6) を洗浄を繰り返します。 注意: 洗濯手順中に軽く動作するように重要です。カテーテルまたは自動の障害で血液流出液を尿道を貫通は通常を示します。エレクトロポレーション効率が低下するにも、気泡を導入することを避けるために重要です。</li>
</ol>4. プラスミド配信
<ol><li>70% エタノールをマウスの腹腔に適用し、膀胱の上腹部の皮膚に 1 cm の縦切開をするメスを使用します。</li>
	<li>、カテーテルの外側の端に少なくとも 20 μ L のプラスミド プラス トリパン ブルー溶液をピペット、注射器を取り付けます。</li>
	<li>膀胱内にプラスミド溶液を注入します。膀胱が青に変わったら、注入は成功します。カテーテルを膀胱内に空気の泡を作成するを避けるためにいくつかの液体を残します。</li>
	<li>ピンセットを使用して外部の尿道口をつかむ、カテーテルと注射器を取り外します。文字列を使用して、外部の尿道口を締めます。文字列に 2 つのループを作るし、2 つのノットで結びます。 注: は、尿道の締付プラスミド液体の逆流を防止するために重要です。</li>
	<li>次のパラメーターでエレクトロポレーション ジェネレーターをオンに: 33 v 系、50 ms の稼働時間、および 950 ms 間隔時間。</li>
	<li>ピンセットで膀胱を押し、2 つの電極と膀胱をピンチします。フットペダル、エレクトロポレーションを実行するを押してください。 注: 各ペダル プッシュ後 5 回短い間隔で発生する電気パルスが設定されます。マウスは、このプロセスでけいれんがあります。配信の効率を増やすことができます足のペダルを 2 回以上押すが、あまりにも多くの電気パルスは尿路の組織の整合性を損なうことができます。<ol>
<li>火花が生成されます、ピンセットと電極との間の接触を避けてください。</li>
	</ol>
</li>
</ol>5. 回復
<ol><li>滅菌手術用縫合糸やクリップの出店を腹部、皮膚を縫合します。傷にヨウ素を適用します。</li>
	<li>イソフルラン システムから動物を削除します。手術後の痛みを軽減するため皮下にブプレノルフィン鎮痛剤 (0.1 mg/kg) を管理します。</li>
	<li>加熱パッドで動物を飼うし、完全復旧後自宅のケージにそれを返します。通常の機動性、飲酒や摂食行動と感染の兆候切開が癒されるし、クリップを削除するまでに、少なくとも一度毎日動物を確認します。動物の痛みの症状を表示は、グルーミング、増加攻撃性と発声を削減など、ブプレノルフィン鎮痛薬の後の注射を管理します。</li>
</ol>

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エレクトロポレーションは細胞膜透過性を高め、遺伝子電子伝達系16のための強力な技術です。電気穿孔法による生活を齧歯動物への遺伝子導入は神経生物学分野17,18,19,20,21によく使用されています。他の多くの器官の潜在的な応用は広範囲にわたって?...

遺伝子組み換えマウス モデル (GEMMs) は、動物の開発、遺伝子機能生体内、および疾患進行1,2メカニズムに関する知識の拡大に重要な役割を演奏されています。生殖 GEMMs は伝統的に 2 つの方法で開発されています。最初の偽妊娠女性に受精卵移植による続く DNA のプラスミッドの前核注入によるトランスジェニック マウスの作成です。他のアウト/ノックアウト ・ ノックイン マウス胚性幹細胞での相同組み換えによる生成とキメラの開発です。興味の特定の組織や細胞の種類の遺伝子のノックアウト条件付きにはには、複数世代の Cre、flox のサイトと遺伝子組み換え遺伝子の繁殖が含まれます。全体のプロセスが高価な目標は、組織に複数の遺伝子を対象とする場合は特に、時間のかかることができます。最近では、クラスター化された空間定期的に短い回文繰り返し (CRISPR) などの遺伝子編集技術はモデリング3、土着の癌組織特異遺伝子変異を誘導するマウスの複数の器官に適用されています。 4,5,6,7または8,9その場で正しい病気の突然変異。これらの研究では、gRNA ベクトル配信通常器官または流体の尾静脈注射のアデノ ウイルスやレンチ ウイルス感染によって達成されました。利便性と癌の伝統的な Cre Lox 基づくマウス モデルと比較してこれらの器官のモデリングの効率、肺と肝臓への CRISPR コンポーネントの配信の相対的な容易さが大幅に向上します。
膀胱尿路上皮はほとんど膀胱腫瘍が発生します。膀胱尿路上皮癌モデリングや遺伝子治療への DNA ベクターの配信は、ウイルス感染の10、11の付着のための障壁として機能する頂尿路上皮表面にグリコサミノグリカン層によって妨げられています。Syn3 などドデシル-β-d-maltoside いくつかの前処理剤は、この障壁を破壊し、アデノ ウイルス感染効率12,13,14を高めるために使用されています。アデノ随伴ウイルス (通気) することができます効果的に膀胱に感染するかどうかは報告されていません。全体的にみて、非ウイルス性に基づいて配信方法が望ましいでしょう。ここでは、尿道カテーテル法, エレクトロポレーションによるマウス尿路への効率的な DNA プラスミド配信のラボで開発手法について述べる.我々 のアプローチは、グリコサミノグリカン レイヤーまたはウイルス産生の化学的前処理を含まない、それ自体が大幅 DNA プラスミドのマウス膀胱尿路への導入を簡略化し、様々 な体内調査を促進する必要があります。膀胱疾患。

<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

遺伝子組み換えマウス モデル (GEMMs) は、癌などの人間の病気の開始そして進行のメカニズムを生物学的知見を明らかに非常に貴重です。遺伝子発現形質転換および条件付きノックアウト マウスが頻繁に使用されているまたは特定のティッシュかセルでアブレーション タイプの生体。ただし、生殖マウス モデルの生成は、時間がかかり、高価にすることができます。遺伝子編集技術及びウイルス感染によってプラスミド DNA を提供することの実現可能性の最近の進歩は、いくつかの臓器の非生殖土着マウス癌モデルの急速な生成を有効にしています。膀胱は、ウイルスベクター尿路を覆うグリコサミノグリカン層の存在のために、アクセスするは難しいされている器官です。ここでは、マウス膀胱尿路上皮の体内に DNA プラスミドの効率的な配信のためのラボで開発された手法について述べる.PCAG GFP プラスミドの膀胱内注入および外科的露出膀胱のエレクトロポレーション、膀胱尿路上皮細胞一過性発現のために具体的には DNA のプラスミッドを配信できることを示す.本手法は過剰発現とマウス膀胱における遺伝子のノックダウンのため迅速かつ便利な方法を提供します、膀胱がんや他の泌尿器科疾患の GEMMs を構築に適用することができます。

膀胱尿路上皮への遺伝子導入の成功を示すためには、エレクトロポレーションに pCAG GFP15プラスミドを使用しました。尿道を通って注入されたプラスミドのトリパン ブルー溶液 20 μ L と 5 の電気パルスを投与します。48 時間後は、マウス膀胱を解剖し、負の制御 (図 1 a) GFP シグナルを示さなかったエレクトロポレーションを受?...

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

マウス膀胱の生体内で尿道カテーテル法, エレクトロポレーションによる尿路上皮細胞への DNA のプラスミッドの配信手法について述べる.膀胱疾患の土着のマウス ・ モデルを生成するための迅速かつ便利な方法を提供しています。

電気穿孔法によるマウス膀胱尿路上皮にプラスミド DNA 配信法

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

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

8.4K ビュー.  University of California, Santa Cruz. マウス膀胱の生体内で尿道カテーテル法, エレクトロポレーションによる尿路上皮細胞への DNA のプラスミッドの配信手法について述べる.膀胱疾患の土着のマウス ・ モデルを生成するための迅速かつ便利な方法を提供しています。

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

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.

マウス子宮内でエレクトロポレーション：制御時空間遺伝子導入

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

神経科学

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.

遺伝子送達研究のための同所膀胱癌モデル

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

医学

Neonatal Pial Surface Electroporation

Over the past several years the pial surface has been identified as a germinal niche of importance during embryonic, perinatal and adult neuro- and gliogenesis, including after injury. However, methods for genetically interrogating these progenitor populations and tracking their lineages had been limited owing to a lack of specificity or time consuming production of viruses. Thus, progress in this region has been relatively slow with only a handful of investigations of this location. Electroporation has been used for over a decade to study neural stem cell properties in the embryo, and more recently in the postnatal brain. Here we describe an efficient, rapid, and simple technique for the genetic manipulation of pial surface progenitors based on an adapted electroporation approach. Pial surface electroporation allows for facile genetic labeling and manipulation of these progenitors, thus representing a time-saving and economical approach for studying these cells.

The pial surface is a unique progenitor zone in the CNS that is receiving increasing attention. Herein, we detail a method for rapid genetic manipulation of this progenitor zone using a modified electroporation method. This procedure can be used for cellular and molecular investigations of cell lineages and signaling pathways involved in cell differentiation and to elucidate the fate and properties of daughter cells.

新生児軟膜表面エレクトロ

Over the past several years the pial surface has been identified as a germinal niche of importance during embryonic, perinatal and adult neuro- and ...

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.

マウス精巣の生体内マイクロインジェクションやエレクトロポレーションで

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

生物学

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.

マウス骨格筋へのプラスミドDNAのエレクトロポレーション

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.

アデノウイルスを介した形質導入を用いた天然膀胱尿路上皮における導入遺伝子の発現

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://jove.unicartagenaproxy.elogim.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.

エレクトロポレーションを介したIn vitro遺伝子導入:蛍光レポータータンパク質をコードするプラスミドを培養細胞に導入するための電気パルス技術

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

生物学的手法

遺伝子導入技術

トランスフェクション

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://jove.unicartagenaproxy.elogim.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.

マウスモデルにおける尿の枯渇と膀胱のすすぎ:マウスの膀胱を空にして洗浄するためのカテーテルベースの手順

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 ...

がん研究

尿路がん

体内研究

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://jove.unicartagenaproxy.elogim.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.

マウス膀胱の尿路上皮へのDNAプラスミドの送達:マウス膀胱の尿路上皮細胞にDNAプラスミドを効率的に送達するためのエレクトロポレーションベースの手順により、遺伝子操作モデルを生成する

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://jove.unicartagenaproxy.elogim.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.

流体力学的腎骨盤注射:マウスモデルの腎臓細胞にDNAを直接送達する技術

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, ...