载体构建及病毒包装服务

载体构建及病毒包装服务

价格: ¥2000 - 10000

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服务名称 :提供载体构建,慢病毒(LV)、腺病毒(ADV)、腺相关病毒AAV、逆转录病毒(RV)包装服务

提供商 :OBiO和元生物

规格 :TU/ml, VG/ml, PFU/mL


载体构建及病毒包装服务
和元生物慢病毒(LV)、腺病毒(ADV)、腺相关病毒(AAV)、逆转录病毒(RV)的生产和指控采取了国际学术界公认的标准流程,所获得的病毒完全可以满足各类实验的使用要求:
慢病毒(和元)的滴度在108~109TU/ml
腺病毒(和元)的滴度在1010~1011pfu/ml
腺相关病毒AAV(和元)的滴度在1012~1013v.g./ml
逆转录病毒(和元)滴度在107~108TU/ml

一、病毒载体及病毒包装介绍
病毒载体是当前生物医学研究中重要的基因转导工具。一方面病毒载体经过改造具有更强的使用安全性和更快的分子克隆速度;另一方面,病毒的感染能力得到了定向的人工进化:治疗靶点筛选者希望得到更快捷,更广谱的转导工具,而基因治疗却需要更安全更特异的感染特性。在不断涌现的新工具新方法面前,研究者常常会无所适从,既难舍对已有系统的信赖与熟悉,又倾心于新工具的蓬勃前景。
目前生物医学研究中常用的病毒载体包括慢病毒,逆转录病毒,腺相关病毒,腺病毒。表1简单的总结了四种病毒载体的基本生物学特性。和元慢病毒感染范围广,表达稳定长效,已经成为分子生物操作中的常规载体,越来越多的分子生物学技术与方法借助于慢病毒载体得以应用。腺病毒载体作为研究和使用时间最长的载体,虽然存在一些不足,但其特点依然鲜明,即感染速度快,基因装载量大。逆转录载体因其特殊的感染特性,仍在干细胞和血液细胞等分裂细胞研究中占有一席之地。而势头强劲的腺相关病毒因其极高的滴度和低转基因安全性,成为基因治疗研究的新宠儿。
表1 不同病毒的生物学特性比较
病毒特性 病毒类型
慢病毒(LV) 逆转录病毒(RV) 腺相关病毒(AAV) 腺病毒(ADV)
包膜
颗粒直径 90-100 nm 90-100 nm 20-30 nm 60-90nm
基因组 dsRNA dsRNA ssDNA dsDNA
表达起始时间 48-72h 48-72h 72-96h 24-48h
表达持续时间 > 2 months > 2 months > 6 months < 1 month
整合方式 随机高频整合 随机高频整合 定向低频整合 非整合

1.腺相关病毒
和元腺相关病毒(adeno-associated virus,AAV)是一类细小病毒,基因组为单链DNA,对分裂细胞和非分裂细胞均具有感染能力。 AAV的血清型是指目前已发现的不同衣壳蛋白的数十种AAV在体内产生不同的抗血清类型,它们对相同的组织和细胞具有不同的感染效率。
重组腺相关病毒rAAV是利用AAV2型基因组与不同的衣壳蛋白结合产生的混合体病毒载体,一般标记为rAAV2/N (N为不同的衣壳血清型)。纽恩生物通过长期的研发,构建了一系列用于基因表达和基因干扰操作的载体。
表2 不同血清型AAV的组织嗜亲性
血清型 适用组织 胞外吸附分子
AAV1 肌肉,心脏,神经 未知
AAV2 体外,神经 硫酸乙酰肝素蛋白多糖
AAV5 肺,眼睛,神经 N-唾液酸
AAV6 肺,心脏 未知
AAV7 肌肉,肝脏 未知
AAV8 肝脏,肌肉,眼睛 未知
AAV9 肺,肝脏,肌肉,心脏 未知
rh10 肋膜,神经 未知

病毒感染实例如下:

2.慢病毒
慢病毒(Lentivirus): 是一类改造自人免疫缺陷病毒(HIV)的病毒载体。它是一种RNA病毒,可利用逆转录酶将外源基因整合到基因组中实现稳定长时表达。目前广泛用于哺乳动物细胞和整体的基因转化操作,具有能同时感染分裂期与非分裂期的细胞等特性。

病毒感染实例如下:

3. 逆转录病毒
逆转录病毒(Retrovirus) :改自鼠白血病病毒(MMLV),是一种RNA病毒,可以利用逆转录酶将外源基因整合到基因组中实现稳定长时表达。逆转录病毒特异性地感染分裂细胞,对造血干细胞和神经干细胞在体外和体内感染具有良好的效果。
病毒感染实例如下:
4. 腺病毒
腺病毒 (Adenovirus) :是一种DNA病毒,具有广泛的细胞和组织感染能力。感染过程剧烈,适于在短时间内进行基因高表达的实验。与其它病毒载体相比,腺病毒最大优势在于插入片段较长,表达活性强;可进行扩增操作。
病毒感染实例如下:

二、客户代表性文章列表
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[11] Nature Communications. (IF=11.329).Wei PF,et.al.(2015).Processing of visually evoked innate fear by a non-canonical thalamic pathway.[腺相关病毒,神经,丘脑]
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[13] Nature Communications. (IF=11.329).Han G,et.al.(2016).Hexose enhances oligonucleotide delivery and exon skipping in dystrophin-deficient mdx mice.[腺相关病毒,肌肉注射,基因治疗]
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[15] Embo Journal. (IF=9.643).Ren SQ,et.al.(2013).PKCλ is critical in AMPA receptor phosphorylation and synaptic incorporation during LTP.[腺相关病毒,神经系统,大脑皮层]
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[62] International Journal of Neuropsychopharmacology. (IF=4.333).Wang YX,et.al.(2014).Protein kinase Mζ is involved in the modulatory effect of fluoxetine on hippocampal neurogenesis in vitro.[腺相关病毒,海马,定位注射]
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[69] Cell Cycle. (IF=3.952).Meng ZS,et.al.(2015).Tumorigenicity Analysis of Heterogeneous Dental Stem Cells and Its Self-Modification for Chromosome Instability.[慢病毒,牙髓干细胞感染]
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[74] Journal of Vascular Surgery. (IF=3.454).Sun Y,et.al.(2016).The regulatory role of smooth muscle 22 on the proliferation of aortic smooth muscle cells participates in the development of aortic dissection.[慢病毒,主动脉平滑肌细胞,干扰]
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[77] Plos One. (IF=3.057).Yang C,et.al.(2013).Systemic Blockade of Dopamine D2-Like Receptors Increases High-Voltage Spindles in the Globus Pallidus and Motor Cortex of Freely Moving Rats.[腺相关病毒,纹状体,定位注射]
[78] Plos One. (IF=3.057).Kuang CY,et.al.(2014).Schlafen 1 Inhibits the Proliferation and Tube Formation of Endothelial Progenitor Cells.[腺病毒,外周血内皮祖细胞,过表达]
[79] Plos One. (IF=3.057).Zeng J,et.al.(2014).GSK3β Overexpression Indicates Poor Prognosis and Its Inhibition Reduces Cell Proliferation and Survival of Non-Small Cell Lung Cancer Cells.[慢病毒,非小细胞肺癌,基因表达]
[80] Plos One. (IF=3.057).Chen Y,et.al.(2016).Increased Expression of SETD7 Promotes Cell Proliferation by Regulating Cell Cycle and Indicates Poor Prognosis in Hepatocellular Carcinoma.[CCK-8]
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