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

Biz Profesör Bin Chen, Dr Yue Zhang ve Kendy Hoang elektroporasyon sistemi kurma konusunda yardım için teşekkür ederim. Bu iş Z.A.W. için Santa Cruz kanser fayda grubu ve NIH gelen hibe tarafından desteklenmiştir

Burada açıklanan tüm yordamları kurallar ve düzenlemeler kurumsal hayvan bakım ve kullanım Komitesi (IACUC) Kaliforniya Üniversitesi Santa Cruz uygun olarak gerçekleştirilmiştir.
1. Plazmid ve araçları hazırlama
<ol><li>Her mesane transfect, DNA plazmid (1 µg/µL) en az 20 µL hazırlamak.</li>
	<li>Trypan mavi 1 µL plazmid çözüm 1.5 mL microcentrifuge tüp içinde 20 µL ekleyin. Pipet iyice karıştırın.</li>
	<li>Otoklav tüm cerrahi aletleri ve çalışma alanı % 70...

<ol>
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W., Chin, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-germline+genetically+engineered+mouse+models+for+translational+cancer+research.">Non-germline genetically engineered mouse models for translational cancer research.</a> Nat Rev Cancer. 10 (7), 470-480 (2010).</li><li>Maddalo, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+engineering+of+oncogenic+chromosomal+rearrangements+with+the+CRISPR/Cas9+system.">In vivo engineering of oncogenic chromosomal rearrangements with the CRISPR/Cas9 system.</a> Nature. 516 (7531), 423-427 (2014).</li><li>Xue, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CRISPR-mediated+direct+mutation+of+cancer+genes+in+the+mouse+liver.">CRISPR-mediated direct mutation of cancer genes in the mouse liver.</a> Nature. , (2014).</li><li>Sanchez-Rivera, F. 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J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CRISPR-Cas9+knockin+mice+for+genome+editing+and+cancer+modeling.">CRISPR-Cas9 knockin mice for genome editing and cancer modeling.</a> Cell. 159 (2), 440-455 (2014).</li><li>Yin, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genome+editing+with+Cas9+in+adult+mice+corrects+a+disease+mutation+and+phenotype.">Genome editing with Cas9 in adult mice corrects a disease mutation and phenotype.</a> Nat Biotechnol. 32 (6), 551-553 (2014).</li><li>Yang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+dual+AAV+system+enables+the+Cas9-mediated+correction+of+a+metabolic+liver+disease+in+newborn+mice.">A dual AAV system enables the Cas9-mediated correction of a metabolic liver disease in newborn mice.</a> Nat Biotechnol. 34 (3), 334-338 (2016).</li><li>Negrete, H. O., Lavelle, J. P., Berg, J., Lewis, S. A., Zeidel, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Permeability+properties+of+the+intact+mammalian+bladder+epithelium.">Permeability properties of the intact mammalian bladder epithelium.</a> Am J Physiol. 271 (4 Pt 2), F886-F894 (1996).</li><li>Lilly, J. D., Parsons, C. 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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electroporation+and+RNA+interference+in+the+rodent+retina+in+vivo+and+in+vitro.">Electroporation and RNA interference in the rodent retina in vivo and in vitro.</a> P Natl Acad Sci USA. 101 (1), 16-22 (2004).</li><li>Yarmush, M. L., Golberg, A., Sersa, G., Kotnik, T., Miklavcic, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electroporation-based+technologies+for+medicine:+principles,+applications,+and+challenges.">Electroporation-based technologies for medicine: principles, applications, and challenges.</a> Annu Rev Biomed Eng. 16, 295-320 (2014).</li><li>Saito, T., Nakatsuji, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+gene+transfer+into+the+embryonic+mouse+brain+using+in+vivo+electroporation.">Efficient gene transfer into the embryonic mouse brain using in vivo electroporation.</a> Dev Biol. 240 (1), 237-246 (2001).</li><li>Boutin, C., Diestel, S., Desoeuvre, A., Tiveron, M. C., Cremer, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+in+vivo+electroporation+of+the+postnatal+rodent+forebrain.">Efficient in vivo electroporation of the postnatal rodent forebrain.</a> PLoS One. 3 (4), e1883 (2008).</li><li>Saito, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+electroporation+in+the+embryonic+mouse+central+nervous+system.">In vivo electroporation in the embryonic mouse central nervous system.</a> Nat Protoc. 1 (3), 1552-1558 (2006).</li><li>Molotkov, D. A., Yukin, A. Y., Afzalov, R. A., Khiroug, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+delivery+to+postnatal+rat+brain+by+non-ventricular+plasmid+injection+and+electroporation.">Gene delivery to postnatal rat brain by non-ventricular plasmid injection and electroporation.</a> J Vis Exp. (43), (2010).</li><li>Walantus, W., Castaneda, D., Elias, L., Kriegstein, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+utero+intraventricular+injection+and+electroporation+of+E15+mouse+embryos.">In utero intraventricular injection and electroporation of E15 mouse embryos.</a> J Vis Exp. (6), e239 (2007).</li><li>Follenzi, A., Santambrogio, L., Annoni, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+responses+to+lentiviral+vectors.">Immune responses to lentiviral vectors.</a> Curr Gene Ther. 7 (5), 306-315 (2007).</li></ol>

Elektroporasyon hücre membran geçirgenliği artırır ve gen electrotransfer16için güçlü bir teknolojidir. Gen teslim yaşam kemirgenler tarafından elektroporasyon içine Nörobiyoloji alanları17,18,19,20,21' sık kullanılan. Onun potansiyel uygulamalar birçok diğer organlarda kapsamlı bir şekilde araştırılmalıdır değ...

Genetiği fare modelleri (GEMMs) hayvan geliştirme, gen fonksiyonları vivo içindeve mekanizmaları hastalık ilerleme1,2hakkında bilgimizi genişletmek önemli bir rol oynuyor. Germline GEMMs geleneksel olarak iki şekilde geliştirilmiştir. İlk DNA plazmid pseudopregnant kadın zigotları nakli tarafından takip pronuclear enjeksiyonla transgenik fareler oluşturulmasıdır. Diğer knock-out/knock-gelen fareler tarafından homolog rekombinasyon embriyonik kök hücre üretimi ve chimeras gelişimi bu. Genlerin ilgi belirli doku veya hücre türü koşullu nakavt Cre ve flox siteleri ile genetik allelleri üreme birden çok nesiller için içerir. Kül oluşum-ebilmek var olmak pahalı ve zaman alıcı, özellikle hedef doku-özellikle birden çok gen hedef için ise. Son zamanlarda, gen düzenleme teknolojileri kümelenmiş düzenli olarak interspaced kısa palindromik yineler (CRISPR) gibi çeşitli organlarda doku özgü genetik değişiklikler otokton kanseri3modelleme için ikna etmek için farelerin uygulanmış olan, 4,5,6,7 veya doğru hastalık mutasyonlar in situ8,9. Bu çalışmalarda gRNA vektörler teslimini genellikle adenoviral veya lentiviral enfeksiyonu organ veya hidrodinamik kuyruk-damar enjeksiyon elde edildi. Akciğer ve karaciğer için CRISPR bileşenlerinin teslimatlar göreli kolaylığı büyük ölçüde kolaylık ve geleneksel Cre-Lox göre fare modellerle karşılaştırıldığında bu organlarda modelleme kanser verimliliği iyileşmiştir.
Mesane urothelium nereden köken çoğu mesane tümörleri var. Mesane urothelium kanser modelleme veya gen tedavisi için DNA vektörel çizimler teslimini glikozaminoglikan katmanı apikal ürotelyal yüzeyinde, bulaşıcı virüs10,11bağlılık için bir engel olarak engel oluyor. Syn3 ve lauryl-β-d-maltoside gibi birkaç tedavi öncesi Aracısı bu bariyer bozabilir ve adenovirus enfeksiyonu verimliliği12,13,14geliştirmek için kullanılmaktadır. Olup olmadığını virüs (AAVs) Adeno ilişkili etkin bir şekilde-ebilmek bulaştırmak mesane bildirilmemiştir. Genel olarak, bir viral tabanlı teslimat yöntemi arzu olurdu. Burada, verimli DNA plazmid teslim edilmek üzere fare urothelium İdrar Kateterizasyon ve elektroporasyon Laboratuarında geliştirilmiş bir roman yöntemi açıklanmaktadır. Yaklaşımımız kimyasal Önarıtma glikozaminoglikan katman veya virüs üretim anlamına gelmez çünkü o önemli ölçüde DNA plazmid teslim fare mesane urothelium içine kolaylaştırır ve çeşitli in vivo çalışmalar kolaylaştırmak gerektiğini Mesane hastalıkları.

<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&#38;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 &#181;L</td>
			<td>USA Scientific</td>
			<td>1111-0206</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipet Tips, Universal Fit; 0.1 to 10 &#181;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&#34; 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

Genetiği fare modelleri (GEMMs) insan hastalıkları kanser gibi başlama ve ilerleme mekanizmalarının içine roman biyolojik anlayışlar ortaya son derece değerlidir. Transgenik ve koşullu nakavt fareler sık gen overexpression için kullanılmış olan veya ablasyon belirli doku veya hücre içinde vivotürleri. Ancak, germline fare modelleri oluşturma zaman alıcı ve pahalı olabilir. Gen teknolojileri ve DNA plazmid tarafından viral enfeksiyon teslim fizibilite düzenleme son gelişmeler hızlı oluşturulmasında çeşitli organları germline sigara otokton fare kanser modelleri etkinleştirdiniz. Mesane urothelium kapsayan bir glikozaminoglikan katman varlığı nedeniyle erişmeye viral vektörler için zor olan bir organdır. Burada, laboratuar DNA plazmid verimli bir şekilde teslim içine fare mesane urothelium içinde vivoiçin geliştirilen bir roman yöntemi açıklanmaktadır. PCAG-GFP DNA plazmid İntravezikal korumak ve cerrahi olarak maruz mesane çoğalmasıyla, DNA plazmid geçici ifade için mesane ürotelyal hücre içine özellikle teslim olduğunu göster. Bizim yöntem overexpression ve fare mesane genlerin nakavt için hızlı ve uygun bir yol sağlar ve GEMMs mesane kanseri ve diğer üroloji hastalıkları bina için uygulanabilir.

Gen teslim mesane urothelium içine başarı göstermek için biz pCAG-GFP15 plazmid elektroporasyon için kullanılır. Plazmid ve Trypan mavi çözüm 20 µL üretra enjekte ettiler ve 5 elektrik darbeleri idare. 48 h sonra fare mesane disseke ve negatif kontrol ise GFP sinyal (şekil 1A) gösterdi elektroporasyon geçmesi değil mesane GFP floresan bir diseksiyon floresan mikroskop altında yamalari gözlendi. Cytokeratin (CK) 5, CK...

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

Biz'DNA plazmid dır fare mesane vivo içinde İdrar Kateterizasyon ve elektroporasyon ürotelyal hücre içine bir roman yöntemi açıklanmaktadır. Kesesi hastalıkları otokton fare modelleri oluşturmak için hızlı ve kolay bir yol sunar.

Plazmid DNA teslimat için fare mesane Urothelium elektroporasyon tarafından roman yöntemi

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

The overall goal of this method is to deliver DNA plasmids into the bladder urothelium of live mice in vivo for gene overexpression or genome editing. This method can help answer key questions in the bladder cancer field, such as how do different genetic mutations promote bladder cancer progression? The main advantage of this technique is that it provides a fast and convenient way for delivering genes into bladder without the safety concerns and technical barriers associated with viral vectors.I first had the idea for this method of trying to use any engineered associated viruses for DNA delivery, which didn't work well the bladder. The implication of this technique can also extend towards gene therapy because electroporation can achieve efficient delivery of DNA plasmids into the urothelial cells. Demonstrating the procedure will be Ofir Stefanson and Yueli Liu from my laboratory.To begin this procedure, autoclave all surgical instruments and sterilize the workspace with 70%ethanol. Spray surgical instruments and the gloves with 70%ethanol frequently when performing the subsequent procedures to maintain sterile conditions. To begin, anesthetize the mouse and shave the surgical area.To deplete the urine, apply lubricant to the catheter and ensure that its outer surface is fully covered. Next, insert the catheter into the urethra opening of the mouse and slowly push it until it reaches the bladder. Gently press the abdomen to help urine depletion.Subsequently, discard the urine with a pipette into a waste beaker, then pipette 80 microliters of PBS into the outer end of the catheter with the other end still in the bladder. Next, carefully attach a one millimeter syringe to the catheter. Gently push the syringe to inject PBS into the bladder.Leave some PBS in the catheter to avoid creating air bubbles in the bladder. Afterward, remove the syringe and wait for PBS to drain out of the bladder. Gently press the abdomen to evacuate PBS, then discard the PBS in the catheter with a pipette into the waste beaker.Repeat PBS washing for two more times. For each bladder to transfect, prepare at least 20 microliters of DNA plasmid at one microgram per microliter. Next, add one microliter of trypan blue to the 20 microliters of plasmid solution and a 1.5 milliliter microcentrifuge tube and pipette to mix well.To deliver the plasmid, apply 70%ethanol to the abdomen. Make a vertical incision of one centimeter to open the abdominal skin above the bladder. Next, cut the muscle layer to expose the bladder.Pipette at least 20 microliters of plasmid plus trypan blue solution into the outer end of the catheter and detach it to the syringe. Then, 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. Afterward, grab the external urethral oraifice and 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. This is a crucial step that requires two people to cooperate. Make sure the knots are tight so the plasmids will not leak out.Now, turn on the electroporation generator. Pinch the bladder with two electrodes and push the foot pedal to perform electroporation. To keep the bladder propped one can pinch the back of the abdominal incision with tweezers.Avoid any contact between tweezers and the electrodes as this may generate a spark. When it is done, suture the abdominal and skin opening with sterile, surgical suture. Remove the string and apply clips.Apply iodine onto the wound, and then remove the animal from the isoflurane system. Administer buprenorphine analgesic subcutaneously to alleviate post surgical pain. 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. Then, a week later, remove the clips. Administer subsequent injections of buprenorphine analgesic if the animal displays pain symptoms such as reduced grooming, increased aggressiveness, and vocalization.To demonstrate successful gene delivery into the bladder urothelium, 20 microliters of pCAG-GFP plasmid and trypan blue solution was injected through the urethra and five electric pulses were administered. Shown here is the successful electroporation that turned the bladder green after 48 hours, whereas a negative control bladder that did not undergo electroporation showed no GFP signal. Immunofluorescent staining of sectioned bladder tissues was performed with CK5, CK18, and GFP antibodies.GFP signal was observed to be present in umbrella, intermediate, and basal cells, but not at the muscle layer indicating good specificity of plasmid delivery into the urothelium. Once mastered, this technique can be performed in 15 minutes if performed properly. After its development, this technique paves the way for researchers in the field of bladder cancer to build new autochthonous mouse models to study disease progression mechanisms.

Research

JoVE Journal

Biology

Purifying Plasmid DNA from Bacterial Colonies Using the Qiagen Miniprep Kit

Plazmid DNA Qiagen Miniprep Kit kullanarak Bakteri kolonileri Arındırıcı

Plasmid DNA purification from E. coli is a core technique for molecular cloning. Small scale purification (miniprep) from less than 5 ml of bacterial ...

Biyoloji

Transformation of Plasmid DNA into E. coli Using the Heat Shock Method

Isı Şoku Yöntemiyle E. coli içine Plazmid DNA Dönüşüm

Transformation of plasmid DNA into E. coli using the heat shock method is a basic technique of molecular biology. It consists of inserting a foreign ...

DNA Transfection of Mammalian Skeletal Muscles using In Vivo Electroporation

DNA Transfection of Mammalian Skeletal Muscles using In Vivo Electroporation

Memeli İskelet Kaslar kullanarak DNA Transfeksiyon In Vivo Elektroporasyon

 A growing interest in cell biology is to express transgenically modified forms of essential proteins (e.g. fluorescently tagged constructs and/or mutant ...

Primary Culture and Plasmid Electroporation of the Murine Organ of Corti. 

İlköğretim Kültür ve Corti murin Organ Plazmid Elektroporasyon.

In all mammals, the sensory epithelium for audition is located along the spiraling organ of Corti that resides within the conch shaped cochlea of the ...

Streamlined Purification of Plasmid DNA From Prokaryotic Cultures

Prokaryotik Kültürler itibaren Plazmid DNA Aerodinamik Arıtma

We describe the complete process of AcroPrep Advance Filter Plates for 96 plasmid preparations, starting from prokaryotic culture and ending with high ...

Direct Delivery of MIF Morpholinos Into the Zebrafish Otocyst by Injection and Electroporation Affects Inner Ear Development

Enjeksiyon ve Elektroporasyon Zebra balığı Otocyst içine MIF Morpholinos Stoktan teslim İç Kulak Geliştirme etkiler

In recent years, electroporation has become a popular technique for in vivo transfection of DNA, RNA, and morpholinos into various tissues, including the ...

A Practical and Novel Method to Extract Genomic DNA from Blood Collection Kits for Plasma Protein Preservation

Plazma Protein Koruma Kan Alma Kitleri Genomik DNA Ayıklama Pratik ve Yeni Bir Yöntem

Laboratory tests can be done on the cellular or  fluid portions of the blood. The use of different blood collection tubes  determines the portion of the ...

A Novel Ex vivo Culture Method for the Embryonic Mouse Heart

A Novel Ex vivo Culture Method for the Embryonic Mouse Heart

Yeni Bir<em&gt; Ex vivo</emEmbriyonik Fare Kalp için&gt; Kültür Yöntemi

Developmental studies in the mouse are hampered  by the inaccessibility of the embryo during gestation. Thus, protocols to  isolate and culture individual ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

Yetişkin Fare Kalvaryal Kemik İliği içinde Hematopoietik kök hücrelerin Vivo 4-Boyutlu Takip

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Plasmid-derived DNA Strand Displacement Gates for Implementing Chemical Reaction Networks

Plazmid kökenli Uygulama Kimyasal Reaksiyon Ağları için DNA Strand Deplasman kapıları

DNA nanotechnology requires large amounts of highly pure DNA as an engineering material. Plasmid DNA could meet this need since it is replicated with high ...