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- 各类病毒比较
- Lentivirus简介
- GeneChem Lentivirus vectors
- GeneChem病毒构建服务
- 对照病毒：GFP标记，表达无Knockdown效应的siRNA
- Reference
- 基于lentivirus shRNA技术资料
- lentiviral vector 和 adenovirial vector对不同肿瘤细胞的感染效率
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>> 各类病毒比较 |
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| 病毒表达系统 |
腺病毒表达系统 |
慢病毒表达系统 |
森林脑炎病毒表达系统 |
| 病毒基因组 |
双链DNA病毒 |
RNA病毒 |
RNA病毒 |
| 复制 |
自主复制 |
自主复制 |
自主复制 |
| 是否整合 |
病毒基因组游离于宿主基因组外，瞬时表达外源基因 |
病毒基因组整合于宿主基因组，长时间、稳定表达外源基因 |
病毒基因组游离于宿主基因组外，瞬时表达外源基因 |
| 感染细胞类型 |
感染分裂和不分裂细胞 |
感染分裂和不分裂细胞 |
感染分裂和非分裂细胞，能特异感染神经元 |
| 表达丰度 |
高水平表达 |
中水平表达 |
高水平表达 |
| 表达时间 |
快（1-2 天） |
慢（2-4 天） |
快（6-8h ） |
| 滴度 |
滴度高达1012pfu/ml |
lenti 最高可达109－10TU/ml |
SFV:1012TU/ml |
| 克隆容量 |
可插入高达8kb的外源片段，滴度随插入片段长度增加而降低 |
可插入不超过8kb的外源片段，滴度随插入片段长度增加而降低 |
可插入不超过5kb的外源片段，滴度随插入片段长度增加而降低 |
| 免疫原性 |
高免疫原性 |
低免疫原性 |
低免疫原性 |
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>> Lentivirus简介 |
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>> GeneChem Lentivirus vectors： |
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Lentivirus RNAi 载体元件构成
| Promoter |
Marker |
用途 |
载体目录号 |
| H1 |
GFP/RFP |
RNA interference |
L01 |
| U6 |
GFP/RFP |
RNA interference |
L02 |
| CMV |
GFP |
Over expression |
L03 |
| Ubiquitin |
GFP |
Over expression |
L04 |
| Non |
GFP |
组织特异性promoter介导的基因表达 |
L05 |
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>> GeneChem病毒构建服务： |
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| 服务类型 |
Ⅰ |
Ⅱ |
Ⅲ |
Ⅳ |
Ⅴ |
Ⅵ |
| 病毒产量（病毒颗粒数） |
>1*107 |
>5*107 |
>1*108 |
>2*108 |
>4*108 |
>1*109 |
| 可提供病毒数量（病毒颗粒数） |
1*107~8 |
5*107~8 |
1*108~9 |
2*108~9 |
4*108~9 |
1*109~10 |
| 浓缩倍数 |
- |
100× |
100× |
1000× |
1000× |
1000× |
| 滴度测定 |
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√ |
√ |
√ |
√ |
√ |
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>> 对照病毒：GFP标记，表达无Knockdown效应的siRNA： |
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·siRNA 对照病毒颗粒
| 产品名称 |
描述 |
目录号 |
| Non-silencing-L |
1×107 TU，1×107～8TU/ml, RNAi 阴性对照，用于细胞学实验 |
L-S-N-C |
| 1×108 TU，1×109TU/ml, RNAi 阴性对照，用于in vivo实验 |
L-S-N-V |
| GAPDH-L |
1×107 TU，1×107～8TU/ml, RNAi 阳性对照，用于细胞学实验 |
L-S-G-C |
| 1×108 TU，1×109TU/ml, RNAi 阳性对照，用于in vivo实验 |
L-S-G-V |
·Over expression 对照病毒颗粒
| 产品名称 |
描述 |
目录号 |
| GFP |
1×106 TU，1×106～7TU/ml,
RNAi阴性对照，用于细胞学实验 |
L-E-N-C |
| 1×108 TU，1×109TU/ml,
RNAi阴性对照，用于in vivo实验 |
L-E-N-V |
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>> Reference： |
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- "Li M, Rossi JJ." Lentiviral Vector Delivery of siRNA and shRNA Encoding Genes into Cultured and Primary Hematopoietic Cells. Methods Mol Biol. 2005;309:261-72.
- "Fish RJ, Kruithof EK." Short-term cytotoxic effects and long-term instability of RNAi delivered using lentiviral vectors. BMC Mol Biol. 2004 Aug 3;5(1):9.
- "Nishitsuji H, Ikeda T, Miyoshi H, Ohashi T, Kannagi M, Masuda T." Expression of small hairpin RNA by lentivirus-based vector confers efficient and stable gene-suppression of HIV-1 on human cells including primary non-dividing cells. Microbes Infect. 2004 Jan;6(1):76-85.
- "An DS, Xie Y, Mao SH, Morizono K, Kung SK, Chen IS." Efficient lentiviral vectors for short hairpin RNA delivery into human cells. Hum Gene Ther.
2003 Aug 10;14(12):1207-12.
- "Scherr M, Battmer K, Ganser A, Eder M." Modulation of gene expression by lentiviral-mediated
delivery of small interfering RNA. Cell Cycle. 2003 May-Jun;2(3):251-7.
- "Scherr M, Battmer K, Dallmann I, Ganser A, Eder M." Inhibition of GM-CSF receptor function by stable RNA interference in a NOD/SCID mouse hematopoietic stem cell transplantation model. Oligonucleotides.
2003;13(5):353-63.
- "Abbas-Terki T, Blanco-Bose W, Deglon N, Pralong W, Aebischer P." Lentiviral-mediated RNA interference. Hum Gene Ther. 2002 Dec 10;13(18):2197-201.
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>> 基于lentivirus shRNA技术资料： |
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·特点：
- 直接包装成为假病毒颗粒,对分裂和非分裂细胞均有感染作用；
- 无需任何转染试剂,操作简洁；
- 实验在手时间短；
- 可以通过简单方式，在短时间内获得稳定表达特定基因shRNA的多种细胞株；
- 可以根据客户需要制备多种标记。
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·适合范围：
- 难于转染细胞的RNAi 研究，特别是神经细胞、悬浮细胞、干细胞；
- 建立稳定表达特定基因shRNA的细胞株；
- In vivo 实验，包括稳定表达细胞株成瘤实验、瘤内注射、局部注射等。
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>> lentiviral vector 和 adenovirial vector对不同肿瘤细胞的感染效率： |
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| Cancer type |
cell line |
transduction efficiency(%) |
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Lentivirus |
Adenovirus |
Bladder
Cancer |
J82 |
80.5 (±1.57) |
62.0 (±0.49) |
| TCCSUP |
66.1 (±1.08) |
37.5 (±0.28) |
| UM-UC-3 |
90.5 (±1.63) |
75.4 (±5.37) |
Bone
Cancer |
MG-63 |
18.9 (±3.70) |
14.6 (±0.55) |
| Saos-2 |
87.5 (±2.84) |
44.3 (±4.33) |
| U-2-OS |
92.4 (±1.26) |
90.5 (±0.55) |
Brain
Cancer |
A172 |
70.6 (±5.22) |
74.6 (±1.35) |
| Hs683 |
94.7 (±0.21) |
32.0 (±0.91) |
| IMR-32 |
21.1 (±0.60) |
87.7 (±0.06) |
| SH-SY5Y |
80.5 (±4.70) |
86.3 (±0.26) |
| TE671 |
90.0 (±0.93) |
75.6 (±0.26) |
| U-87 MG |
94.9 (±0.21) |
74.9 (±0.26) |
| U-118-MG |
92.2 (±1.09) |
47.8 (±5.14) |
| U-251 MG |
96.2 (±0.18) |
95.1 (±0.26) |
Breast
Cancer |
MCF-7 |
68.1 (±6.22) |
59.6 (±3.12) |
| SK-BR-3 |
72.6 (±1.62) |
94.6 (±0.87) |
| ZR-75-1 |
41.7 (±3.05) |
67.1 (±1.52) |
Cervix
Cancer |
HeLa |
83.5 (±0.75) |
94.4 (±0.20) |
| SiHa |
81.5 (±2.80) |
95.4 (±0.16) |
Colon
Cancer |
CaCo-2 |
8.8 (±0.57) |
87.8 (±1.81) |
| HCT116 |
90.7 (±1.62) |
90.7 (±0.78) |
| HT-29 |
51.5 (±3.92) |
87.2 (±1.00) |
Lung
Cancera |
A549 (ac) |
91.6 (±0.01) |
75.9 (±0.01) |
| Calu-3 (ac) |
61.9 (±0.04) |
78.9 (±0.02) |
| NCI-H23 (ac) |
72.7 (±0.07) |
87.8 (±0.04) |
| NCI-H82 (sc) |
27.7 (±0.03) |
95.0 (±0.00) |
| NCI-H146 (sc) |
44.4 (±0.03) |
91.1 (±) |
| NCI-H460 (lc) |
51.0 (±0.04) |
57.2 (±0.02) |
| NCI-H520 (sq) |
56.3(±0.07) |
76.3 (±0.00) |
| NCI-H661 (lc) |
75.2 (±0.02) |
80.0 (±0.01) |
| NCI-H1650 (ac) |
81.2 (±0.02) |
80.2 (±0.01) |
| SW-900 (sq) |
54.1 (±0.03) |
80.7 (±0.02) |
| Melanoma |
A2058 |
81.4 (±3.48) |
87.0 (±0.58) |
| SK-MEL-5 |
83.8 (±0.69) |
72.2 (±1.91) |
| WM-115 |
93.6 (±0.38) |
55.4 (±1.52) |
| WM-266-4 |
86.7 (±2.19) |
16.8 (±1.22) |
Pancreatic
cancer |
AsPC-1 |
28.9 (±3.11) |
58.5 (±1.13) |
| BxPC-3 |
48.0 (±0.24) |
58.4 (±2.71) |
| MiaPaCa-2 |
88.4 (±1.41) |
63.2 (±4.86) |
Prostate
Cancer |
22Rv1 |
93.6 (±0.88) |
93.2 (±0.17) |
| DU145 |
47.2 (±1.52) |
92.7 (±0.86) |
| PC-3 |
68.4 (±2.30) |
51.3 (±0.77) |
| 文献来源：INTERNATIONAL JOURNAL OF ONCOLOGY 25: 1753-1762, 2004 |
| In vitro transduction of human cancer cell
lines by lentiviral vector (MOI 3) and adenovirus(MOI 3), as determined
by percentage of GFP positive cells 4 days post-transduction. a: Lung
cancer cell lines are divided into four subtypes: adenocarcinoma (ac),
small cell carcinoma (sc), large cell carcinoma (lc),and squamous cell
carcinoma (sq). |
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化学合成siRNA
