慢病毒

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病毒类型

• VSV-G假病毒化的第三代慢病毒（默认）
• VSV-G假病毒化的第二代慢病毒
• 非整合慢病毒
• 使用其他膜蛋白进行假病毒化的慢病毒（如使用冠状病毒刺突蛋白）
• 无膜蛋白的表面光秃的慢病毒（可用于转导时的阴性对照）

交付内容

对照病毒

为了更好地开展您的实验，我们按照“对照病毒与定制病毒的生物学应用相匹配”的原则给您赠送或者推荐对照病毒。例如，如果定制病毒表达一个基因，则推荐的对照病毒表达荧光蛋白EGFP。如果定制病毒表达打靶某个基因的shRNA，则对照病毒表达一个Scramble shRNA。推荐的对照病毒信息如下表所示：

辅助病毒

为了更好地开展您的实验，我们按照“辅助病毒与定制病毒的生物学应用相匹配”的原则给您推荐辅助病毒。例如，如果定制病毒是一个只含TRE启动子的TetOn慢病毒，则需要一个表达rtTA的辅助病毒，才能进行诱导表达实验。推荐的辅助病毒信息如下表所示：

运输/储存

我们的慢病毒存储在HBSS缓冲液中，并采用干冰运输。收到慢病毒后，立即放置于-80℃，长期贮存至少6个月。慢病毒的保质期约为一年。请避免反复冻融，因为这会导致慢病毒滴度大幅下降。

生产/质控

注意：第三代慢病毒包装系统只能包装第三代的慢病毒转移载体。但是第二代慢病毒包装系统既能包装第二代，也能包装第三代的慢病毒转移载体。第三代慢病毒转移载体上的5’ LTR中的U3元件是被一个外源启动子替代（如CMV启动子或者RSV启动子），使其不依赖Tat蛋白进行HIV RNA基因组的转录，故第三代慢病毒包装系统的包装质粒不含Tat蛋白。第二代慢病毒转移载体上的5’ LTR是野生型的LTR，含有U3、R和U5元件，依赖Tat蛋白转录HIV RNA基因组，故第二代慢病毒包装系统的包装质粒含有Tat蛋白。

滴度保证

我们的滴度保证仅适用于这样的转移载体

• 载体上的慢病毒基因组（从5’ LTR的R元件的第一个碱基到3’ LTR的R元件的最后一个碱基）长度小于承载限度长度9.2 kb（～野生型HIV-1基因组长度）。超过该范围，载体上的病毒基因组也许会被包装进病毒，但会造成病毒滴度降低。

对于如下这样的转移载体，我们不能保证滴度：

• 包含对包装过程有不利影响的序列，例如毒性基因（如促凋亡基因、针对HIV的host factors）；破坏包装细胞完整性的基因（如导致细胞聚集的膜蛋白）、破坏慢病毒RNA基因组完整性的序列（如含有polyA信号序列）、易于重排或者产生二级结构的序列（如重复序列或者高GC含量序列）。
• 用户提供的载体，因为我们无法控制载体质量。

交付周期

交付周期是指订单接收到发货之间的时间，不包括需要客户提供任何物料（如病毒载体）的等待时间。对于不含有荧光标记，但是含有药筛标记的病毒载体，如果您需要获得细胞药筛效果的质检结果，由于细胞生长状态需要较长时间观察，周期将额外增加8-10天。

客户提供载体

如需客户提供慢病毒载体，请联系我们了解外来物品接收指南。请严格按照我们的指南来进行装运以免造成物品的任何延误和损坏。客户提供的所有材料均需经过QC检测，每一份材料附加检测费用。请注意，客户提供物品接收并通过QC之后生产才正式开始。

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慢病毒转导

文档

使用说明书

慢病毒体内应用

COA

常见问题解答

精选文献

FALCON systematically interrogates free fatty acid biology and identifies a novel mediator of lipotoxicity

Cell Metabolism, 2023, doi: 10.1016/j.cmet.2023.03.018

使用的云舟生物产品或服务：

• Lentivirus Packaging
• Vector Cloning

Abstract: Cellular exposure to free fatty acids (FFAs) is implicated in the pathogenesis of obesity-associated diseases. However, there are no scalable approaches to comprehensively assess the diverse FFAs circulating in human plasma. Furthermore, assessing how FFA-mediated processes interact with genetic risk for disease remains elusive. Here, we report the design and implementation of fatty acid library for comprehensive ontologies (FALCON), an unbiased, scalable, and multimodal interrogation of 61 structurally diverse FFAs. We identified a subset of lipotoxic monounsaturated fatty acids associated with decreased membrane fluidity. Furthermore, we prioritized genes that reflect the combined effects of harmful FFA exposure and genetic risk for type 2 diabetes (T2D). We found that c-MAF-inducing protein (CMIP) protects cells from FFA exposure by modulating Akt signaling. In sum, FALCON empowers the study of fundamental FFA biology and offers an integrative approach to identify much needed targets for diverse diseases associated with disordered FFA metabolism.

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Identification of natural compounds as SARS-CoV-2 entry inhibitors by molecular docking-based virtual screening with bio-layer interferometry

Dingqi Zhang, et al.

Pharmacological Research, 2021, doi: 10.1016/j.phrs.2021.105820

使用的云舟生物产品或服务：

• Lentivirus Packaging
• Lentivirus Pseudotyped with Coronavirus Spike (S) Protein
• Vector Cloning

Abstract: Coronavirus Disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), which enter the host cells through the interaction between its receptor binding domain (RBD) of spike glycoprotein with angiotensin-converting enzyme 2 (ACE2) receptor on the plasma membrane of host cell. Neutralizing antibodies and peptide binders of RBD can block viral infection, however, the concern of accessibility and affordability of viral infection inhibitors has been raised. Here, we report the identification of natural compounds as potential SARS-CoV-2 entry inhibitors using the molecular docking-based virtual screening coupled with bilayer interferometry (BLI). From a library of 1871 natural compounds, epigallocatechin gallate (EGCG), 20(R)-ginsenoside Rg3 (RRg3), 20(S)-ginsenoside Rg3 (SRg3), isobavachalcone (Ibvc), isochlorogenic A (IscA) and bakuchiol (Bkc) effectively inhibited pseudovirus entry at concentrations up to 100 μM. Among these compounds, four compounds, EGCG, Ibvc, salvianolic acid A (SalA), and isoliensinine (Isl), were effective in inhibiting SARS-CoV-2-induced cytopathic effect and plaque formation in Vero E6 cells. The EGCG was further validated with no observable animal toxicity and certain antiviral effect against SARS-CoV-2 pseudovirus mutants (D614G, N501Y, N439K & Y453F). Interestingly, EGCG, Bkc and Ibvc bind to ACE2 receptor in BLI assay, suggesting a dual binding to RBD and ACE2. Current findings shed some insight into identifications and validations of SARS-CoV-2 entry inhibitors from natural compounds.

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Glioblastoma remodelling of human neural circuits decreases survival

Saritha Krishna, et al.

Nature, 2023, doi: 10.1038/s41586-023-06036-1

使用的云舟生物产品或服务：

• Lentivirus Packaging
• Vector Cloning

Abstract: Gliomas synaptically integrate into neural circuits. Previous research has demonstrated bidirectional interactions between neurons and glioma cells, with neuronal activity driving glioma growth and gliomas increasing neuronal excitability. Here we sought to determine how glioma-induced neuronal changes infuence neural circuits underlying cognition and whether these interactions infuence patient survival. Using intracranial brain recordings during lexical retrieval language tasks in awake humans together with site-specifc tumour tissue biopsies and cell biology experiments, we fnd that gliomas remodel functional neural circuitry such that task-relevant neural responses activate tumour-infltrated cortex well beyond the cortical regions that are normally recruited in the healthy brain. Site-directed biopsies from regions within the tumour that exhibit high functional connectivity between the tumour and the rest of the brain are enriched for a glioblastoma subpopulation that exhibits a distinct synaptogenic and neuronotrophic phenotype. Tumour cells from functionally connected regions secrete the synaptogenic factor thrombospondin-1, which contributes to the diferential neuron–glioma interactions observed in functionally connected tumour regions compared with tumour regions with less functional connectivity. Pharmacological inhibition of thrombospondin-1 using the FDA-approved drug gabapentin decreases glioblastoma proliferation. The degree of functional connectivity between glioblastoma and the normal brain negatively afects both patient survival and performance in language tasks. These data demonstrate that high-grade gliomas functionally remodel neural circuits in the human brain, which both promotes tumour progression and impairs cognition.

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