Name: intracranial injection of adeno-associated viral vectors
Uploaded: 2010-11-17T18:00:00
Description: Watch this Scientific Journal Video about Intracranial Injection of Adeno-associated Viral Vectors at JoVE.com

这项工作成为可能，从美国国立卫生研究院（EY012977），在宝来惠康基金，白厅基金会，和斯隆基金会（AKM）的生物医学科学事业奖助学金。

1。病毒处理和存储<ol><li>要使用的病毒的生物安全水平的基础上，应选择适当的保护和处理技术。这些做法，可以发现在生物安全微生物和生物医学实验室第 5版，疾病预防控制中心网站上提供 （ <a href="http://www.cdc.gov/od/OHS/biosfty/bmbl5/bmbl5toc.htm">http://www.cdc.gov/od/OHS/biosfty/bmbl5/bmbl5toc.htm</a> ）。 AAV载体的使用已被批准用于1级生物安全（BSL - 1）。实验显示，一个白大褂和手套将处理病?...

<ol>
	<li>Kaplitt, M. G., Leone, P., Samulski, R. J., Xiao, X., Pfaff, D. W., O'Malley, K. L., During, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+gene+expression+and+phenotypic+correction+using+adeno-associated+virus+vectors+in+the+mammalian+brain.">Long-term gene expression and phenotypic correction using adeno-associated virus vectors in the mammalian brain.</a> Nat Genet. 8, 148-154 (1994).</li><li>Harding, T. C., Dickinson, P. J., Roberts, B. N., Yendluri, S., Gonzalez-Edick, M., Lecouteur, R. A., Jooss, K. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhanced+gene+transfer+efficiency+in+the+murine+striatum+and+an+orthotopic+glioblastoma+tumor+model,+using+AAV-7-+and+AAV-8-pseudotyped+vectors.">Enhanced gene transfer efficiency in the murine striatum and an orthotopic glioblastoma tumor model, using AAV-7- and AAV-8-pseudotyped vectors.</a> Hum Gene Ther. 17, 807-820 (2006).</li><li>Lo, W. D., Qu, G., Sferra, T. J., Clark, R., Chen, R., Johnson, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adeno-associated+virus-mediated+gene+transfer+to+the+brain:+duration+and+modulation+of+expression.">Adeno-associated virus-mediated gene transfer to the brain: duration and modulation of expression.</a> Hum Gene Ther. 10, 201-213 (1999).</li><li>Lowery, R. L., Zhang, Y., Kelly, E. A., Lamantia, C. E., Harvey, B. K., Majewska, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+long-term+labeling+of+cells+in+the+developing+and+adult+rodent+visual+cortex+using+double-stranded+adeno-associated+viral+vectors.">Rapid long-term labeling of cells in the developing and adult rodent visual cortex using double-stranded adeno-associated viral vectors.</a> Dev Neurobiol. 69, 674-688 (2009).</li></ol>

维拉利介导的基因传递持有神经过程的研究和治疗脑部疾病1,2,3的巨大潜力。这项技术的伟大的通用性，也可以利用荧光标记细胞在体外和体内 4成像。在这里，我们展示了一个使用双链的表达增强型绿色荧光蛋白的腺协会病毒在小鼠的视觉皮层的神经元和神经胶质细胞的转导的详细过程。 虽然这项技术是比较直截了当的，也有一些需要考虑的技...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>Stoelting Mouse and Neonatal Rat Adaptor</td>
<td>&nbsp;</td>
<td>Stoelting</td>
<td>51625</td>
<td>Regular stereotax for securing animals for surgery may be substituted</td>
</tr>
<tr>
<td>Extra Fine Bonn Scissors, 8.5cm, straight tip, cutting edge 13mm</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14084-08</td>
<td>&nbsp;</td>
<tr>
<td>Eye Dressing Forceps, 10cm, tip width 0.5mm, curved</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11152-10</td>
<td>&nbsp;</td>
<tr>
<td>Dumont #5/45 Forceps- Dumoxel Standard Tip, 11cm, angled</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11251-35</td>
<td>Extra-fine tipped forceps for performing craniotomy</td>
</tr>
<tr>
<td>Standard Pattern Forceps, straight, 2.5mmx1.35mmtip, 12cm</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11000-12</td>
<td>&nbsp;</td>
<tr>
<td>Microtorque Control Box and Tech2000 handpiece</td>
<td>&nbsp;</td>
<td>Ram Products, Inc.</td>
<td>TECH2000ON/OFF</td>
<td>Dental drill</td>
</tr>
<tr>
<td>Micro Drill Stainless Steel Burrs 1.4mm tip diameter</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>19008-14</td>
<td>&nbsp;</td>
<tr>
<td>Wiretrol micropipettes, to deliver 1-5 Ul</td>
<td>&nbsp;</td>
<td>VWR International/ Drummond</td>
<td>5-000-1001 or 53480-287</td>
<td>&nbsp;</td>
<tr>
<td>Mineral oil</td>
<td>&nbsp;</td>
<td>VWR International</td>
<td>29447-338</td>
<td>&nbsp;</td>
<tr>
<td>Manual Micromanipulator and Tilting Base (right-handed)</td>
<td>&nbsp;</td>
<td>World Precision Instruments, Inc.</td>
<td>M3301-M3-R</td>
<td>Used for determining stereotaxic co-ordinates</td>
</tr>
<tr>
<td>UltraMicroPump (UMP3) (one) with SYS-Micro4 Controller</td>
<td>&nbsp;</td>
<td>World Precision Instruments, Inc.</td>
<td>UMP3-1</td>
<td>&nbsp;</td>
<tr>
<td>Sutures</td>
<td>&nbsp;</td>
<td>VWR International</td>
<td>95056-952</td>
<td>&nbsp;</td>
<tr>
<td>P-97 Flaming/Brown Micropipette Puller</td>
<td>&nbsp;</td>
<td>Sutter Instruments</td>
<td>P-97</td>
<td>&nbsp;</td>
<tr>
<td>Tobradex</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Available from your institution&#x2019;s veterinary services</td>
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intracranial injection of adeno-associated viral vectors

颅内的设计，以表达荧光蛋白的病毒载体注射液是一种在体内和不同脑区的脑切片中特定的细胞亚群的可视化的通用标签技术。与荧光染料注射，病毒标签提供针对个别的细胞类型和更便宜和时间消耗比建立转基因小鼠系。在这种技术中，腺相关病毒（AAV）载体注入颅内使用与周围组织的损伤最小立体坐标，一个微管和一个精准投放的AAV的自动泵所需的区域。注射参数，可以根据个人的实验，通过调整注射，注射部位，注射量，注射率，腺相关病毒血清型和基因表达的启动子驾驶的动物年龄。根据选择的条件，可以让病毒诱导的基因表达组细胞，单个细胞或精细胞的过程中，可视化的树突棘水平。这里显示的实验中，描绘了一个双链标签在鼠标的初级视觉皮层的神经元和神经胶质细胞表达绿色荧光蛋白的腺相关病毒注射。

Watch this Scientific Journal Video about Intracranial Injection of Adeno-associated Viral Vectors at JoVE.com

Intracranial Injection of Adeno-associated Viral Vectors

在这里，我们目前颅内注射AAV载体在视觉皮层神经元和神经胶质细胞的荧光标记。

颅内注射腺相关病毒载体

Intracranial injection of viral vectors engineered to express a fluorescent protein is a versatile labeling technique for visualization of specific ...

Research

JoVE Journal

Neuroscience

Targeting of Deep Brain Structures with Microinjections for Delivery of Drugs, Viral Vectors, or Cell Transplants

定向脑深部结构与配送药品，病毒载体，或细胞移植Microinjections

Microinjections into the brain parenchyma are important procedures to deliver drugs, viral vectors or cell transplants.  The brain lesion that an ...

科研

神经科学

Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium

病毒载体的基因治疗的小鼠视网膜色素上皮缘方针，视网膜下注射

The eye is a small and enclosed organ which makes it an ideal target for gene therapy. Recently various strategies have been applied to gene therapy in ...

Intracerebroventricular and Intravascular Injection of Viral Particles and Fluorescent Microbeads into the Neonatal Brain

病毒颗粒和荧光微球侧脑室和血管内注入新生儿脑

In the study on the pathogenesis of viral encephalitis, the infection method is critical. The first of the two main infectious routes to the brain is the ...

Subpial Adeno-associated Virus 9 (AAV9) Vector Delivery in Adult Mice

Subpial Adeno-associated Virus 9 AAV9 Vector Delivery in Adult Mice

在成年小鼠中的亚型腺相关病毒9（AAV9）载体递送

The successful development of a subpial adeno-associated virus 9 (AAV9) vector delivery technique in adult rats and pigs has been reported on previously. ...

Methods and Tips for Intravenous Administration of Adeno-associated Virus to Rats and Evaluation of Central Nervous System Transduction

腺相关病毒对大鼠的静脉管理方法和技巧及中枢神经系统转导的评价

Adeno-associated virus (AAV) vectors are a key reagent in the neurosciences for clustered regularly interspaced short palindromic repeats (CRISPR), ...

Adeno-associated Virus-mediated Transgene Expression in Genetically Defined Neurons of the Spinal Cord

腺相关病毒介导的脊髓基因神经元的转基因表达

Selective manipulation of spinal neuronal subpopulations has mainly been achieved by two different methods: 1) Intersectional genetics, whereby double or ...

Direct Intrathecal Injection of Recombinant Adeno-associated Viruses in Adult Mice

重组腺相关病毒在成年小鼠中的直接注射

Intrathecal (IT) injection of adeno-associated virus (AAV) has drawn considerable interest in CNS gene therapy by virtue of its safety, noninvasiveness, ...

Production, Purification, and Quality Control for Adeno-associated Virus-based Vectors

与腺相关病毒的向量的生产、纯化和质量控制

Gene delivery tools based on adeno-associated viruses (AAVs) are a popular choice for the delivery of transgenes to the central nervous system (CNS), ...

Convection Enhanced Delivery of Optogenetic Adeno-associated Viral Vector to the Cortex of Rhesus Macaque Under Guidance of Online MRI Images

在在线 MRI 图像的指导下,对流增强将视遗传阿德诺相关病毒载体输送到 Rhesus Macaque 的皮质

In non-human primate (NHP) optogenetics, infecting large cortical areas with viral vectors is often a difficult and time-consuming task. Here, we ...

In Vivo Direct Reprogramming of Resident Glial Cells into Interneurons by Intracerebral Injection of Viral Vectors

在Vivo直接重新编程居民胶质细胞到内神经通过病毒载体的脑内注射

Converting resident glia in the brain into functional and subtype-specific neurons in vivo provides a step forward towards the development of alternative ...