【基因不朽化】Gene Immortalization-Myc, p53 Rb Ras CDK4 BmiI Ad5 E1A-E1B, HOX
哺乳動物細胞的不朽化Immortalized策略
存在用於在培養物中不朽化哺乳動物細胞的幾種方法。 Abmgood多年研發經驗，開發出了一系列Immortalized商品；包括即用型逆轉錄病毒，慢病毒和腺病毒載體。細胞不朽化的工具中，SV40 T antigen是最簡單且最常用來造成不朽化細胞生成的方法。多數的病毒基因和抑癌基因（p53、Rb等）的不活化相關，會誘導細胞處於replicative senescence的階段。目前也有研究顯示被感染的細胞，SV40 T anbtigen會誘導端粒酶（Telomerase）的活性。與端粒（telomere）長度相關的Telomerase Reverse Transcriptase protein (TERT)是一種不朽細胞的研究應用。平常細胞中，hTERT是處於不活化態，必要時會進行活化保持telomere的長度，避免細胞發生replicative senescence的現象。某些細胞hTERT 過度表現（over-expression）對於誘發細胞不朽化是無效的（如初代上皮細胞），反而會造成細胞死亡；同時表現hTERT catalytic subunit以及p53或RB siRNA在人類初代卵巢上皮細胞會造成細胞不朽化。過度表現Ras或Myc T58A mutants也有初代細胞不朽化的現象產生。最近的研究發現，hTERT催化亞基與p53或Rb siRNA的共表達可使人，原代，卵巢或上皮細胞永生化，從而提供具有明確定義的遺傳背景的更真實的細胞模型（Yang G. et al。 ，Carcinogenesis 2007; Yang G.等，Oncogene 2007）。類似地，已顯示Cdk4與hTERT的表達使支氣管上皮細胞永生化，所述支氣管上皮細胞既不能單獨永生化（Ramirez R.等，Cell＆Tumor Bio.2004）。Ras V12或Myc T58A突變體的過表達能夠使一些原代細胞類型永生化，並且Bmi-1的過表達可用於使鼻咽和乳腺上皮細胞永生化（Sears R.等，Genes＆Dev。 2000; Song，L。等，Cell，Tumor，＆Stem Cell Bio.2006; Dimri，G。等，Mol.Bio。＆Gen.2002）。HOX基因（HOXB8，HOXA9和HOXA10）是與惡性腫瘤相關的同源域轉錄因子家族，例如急性髓性白血病和急性淋巴性白血病。因此，HOX基因的過表達已被用於永生化各種造血細胞，包括巨噬細胞，造血祖細胞和骨髓祖細胞。人類腺病毒5型的E1A基因作為原型病毒致癌基因已被廣泛研究。該基因已被用於使許多類型的原代囓齒動物細胞永生化，包括腎細胞，上皮細胞，羊水細胞和成纖維細胞。雖然E1A可以自身永生化細胞，但E1B的添加可能通過抑制細胞凋亡來幫助永生化過程。
| Gene Overexpressed | Viruses Offered | Application |
| hTERT | Adenovirus, Lentivirus, Retrovirus |
可靠和常用。 可能在一些細胞類型（例如上皮細胞）中引起細胞死亡 |
| SV40 T Antigen | Adenovirus, Lentivirus, Retrovirus |
可靠和常用。 通常用於不朽化上皮細胞。 |
| EBV genes | Epstein-Barr Virus | 不朽化B淋巴細胞，有時是T淋巴細胞。 |
| HPV E6-E7 | Lentivirus | 不朽化角化細胞。 |
| HOXB8, HOXA9, HOXA10 | Lentivirus | 不朽化造血細胞，包括巨噬細胞，造血祖細胞和骨髓祖細胞。 |
| Adenoviral E1A-E1B | Lentivirus | 不朽化大範圍的初級囓齒動物細胞，包括羊水細胞，原發性上皮細胞，成纖維細胞和原代腎細胞。 |
| p53, Rb siRNA | Lentivirus | 與hTERT共表達以更好地使上皮細胞不朽化。 |
| Bmi1 | Lentivirus | 不朽化上皮細胞。 |
| CDK4 | Lentivirus | 與hTERT共同表達更真實的細胞模型。 |
| Myc T58A (c-Myc), Ras V12 (H-Ras) | Lentivirus | 不朽化各種細胞，包括神經幹細胞，初代腎細胞，成紅細胞和各種上皮細胞（原發性前列腺，乳腺，氣道等）。 |
Abmgood不朽化系列商品 (immortalized reagenet) Ready-to-use
Abmgood公司於人類與老鼠不朽之細胞株上，具有幾十年的實驗經驗；提供一系列不朽 (immortalized)化之細胞株、及不朽化之試劑商品。ready-to-use定量好之retrovirus、lentiviru、 adenovirus，可直接加於細胞中進行感染表現；病毒液包括有SV40重組病毒、EBV重組病毒、hTERT病毒液、Myc基因病毒液、p53基因病毒液、Rb基因病毒液、Ras基因病毒液。提供106及109兩種包裝力價之病毒液；亦提供一系列病毒力價濃縮與定量試劑商品。
細胞不朽化的工具中，SV40 T antigen是最簡單且最常用來造成不朽化細胞生成的方法。多數的病毒基因和抑癌基因（p53、Rb等）的不活化相關，會誘導細胞處於replicative senescence的階段。目前也有研究顯示被感染的細胞，SV40 T anbtigen會誘導端粒酶（Telomerase）的活性。與端粒（telomere）長度相關的Telomerase Reverse Transcriptase protein (TERT)是一種不朽細胞的研究應用。平常細胞中，hTERT是處於不活化態，必要時會進行活化保持telomere的長度，避免細胞發生replicative senescence的現象。某些細胞hTERT 過度表現（over-expression）對於誘發細胞不朽化是無效的（如初代上皮細胞），反而會造成細胞死亡；同時表現hTERT catalytic subunit以及p53或RB siRNA在人類初代卵巢上皮細胞會造成細胞不朽化。過度表現Ras或Myc T58A mutants也有初代細胞不朽化的現象產生。
ABM提供以下不朽細胞的研究工具：
- Gene Immortalization (Myc, p53, Rb, Ras, CDK4, BmiI, Ad5 E1A-E1B, HOX genes) Reagents
- HPV E6-E7 Cell Immortalization Reagents
- SV40 T antigen Cell Immortalization Reagents
- EBV Cell Immortalization Reagents
- hTERT Cell Immortalization Reagents
SV40 Cell Immortalization Reagents不朽化系列商品
HPV Cell Immortalization Reagents不朽化系列商品
| Lenti-HPV-16 E6/E7 Virus | pLenti-III-HPV-16 E6/E7 | Lentivirus containing HPV-16 E6/E7 gene (106 IU/ml) | 10ml | G268 |
| Lenti-HPV-16 E6/E7 Virus, High Titer | pLenti-III-HPV-16 E6/E7 | High Titer Lentivirus containing HPV-16 E6/E7 gene (109IU/ml) | 5 x 20 μl | LV617 |
EBV Cell Immortalization Reagents不朽化系列商品
| Product |
Vector Map (Click to View) |
Description | Quantity | Cat. No. |
| EBV Virus | N/A | Virus containing EBV gene (106 IU/ml) | 10ml | G229 |
hTERT Cell Immortalization Reagents不朽化系列商品
| Product |
Vector Map (Click to View) |
Description | Quantity | Cat. No. |
| Adeno-hTERT | pAdeno-hTERT | Adenovirus containing hTERT (106 IU/ml) | 250 μl | G205 |
| Adeno-hTERT Antisense | pAdeno-hTERT Antisense | Adenovirus containing antisense hTERT (106 IU/ml) | 250 μl | G208 |
| Lenti-CMV-hTERT-GFP-2A-Puro Virus | pLenti-CMV-hTERT-GFP-2A-Puro | Lentivirus containing hTERT with a GFP Reporter & resistance to Puromycin (106 IU/ml) | 10ml | LV623 |
| Lenti-CMV-hTERT-GFP-2A-Puro Virus, High Titer | pLenti-CMV-hTERT-GFP-2A-Puro | High Titer Lentivirus containing hTERT with a GFP Reporter & resistance to Puromycin (109 IU/ml) | 5 x 20 μl | LV624 |
| Lenti-CMV-hTERT-RFP-2A-Puro Virus | pLenti-CMV-hTERT-RFP-2A-Puro | Lentivirus containing hTERT with a RFP Reporter & resistance to Puromycin (106 IU/ml) | 10ml | LV625 |
| Lenti-CMV-hTERT-RFP-2A-Puro Virus, High Titer | pLenti-CMV-hTERT-RFP-2A-Puro | High Titer Lentivirus containing hTERT with a RFP Reporter & resistance to Puromycin (109 IU/ml) | 5 x 20 μl | LV626 |
| Lenti-EF1α-hTERT Virus | pLenti-EF1a-hTERT | Lentivirus containing hTERT (106 IU/ml) | 10ml | G706 |
| Lenti-EF1α-hTERT Virus, High Titer | pLenti-EF1a-hTERT | High Titer Lentivirus containing hTERT (109 IU/ml) | 5 x 20 μl | LV616 |
| Lenti-hTERT virus | pLenti-hTERT | Lentivirus containing hTERT (106 IU/ml) | 10ml | G200 |
| Lenti-hTERT Virus, High Titer | pLenti-hTERT | High Titer Lentivirus containing hTERT (109 IU/ml) | 5 x 20 μl | LV615 |
| Lenti-hTERT Antisense virus | pLenti-hTERT Antisense | Lentivirus containing antisense hTERT (106 IU/ml) | 10ml | G201 |
| Lenti-hTERT-Neo Virus | pLenti-hTERT-Neo | Lentivirus containing hTERT with resistance to Neomycin (106 IU/ml) | 10ml | G204 |
| Lenti-hTERT-Neo Virus, High Titer | pLenti-hTERT-Neo | High Titer Lentivirus containing hTERT with resistance to Neomycin (109 IU/ml) | 5 x 20 μl | LV622 |
| Retro-E1/hTERT virus | pRetro-E1 hTERT | Retrovirus containing hTERT (106 IU/ml) | 10ml | G207 |
Gene Immortalization (Myc, p53, Rb, Ras, CDK4, BmiI, Ad5 E1A-E1B, HOX genes) Reagents基因不朽化商品
| Product |
Vector Map (Click to View) |
Description | Quantity | Cat. No. |
| Lenti-p53 siRNA Virus | Lenti-p53 siRNA | Recombinant p53 siRNA Lentivirus (106 IU/ml) | 10 ml | G219 |
| Lenti-p53 siRNA Virus, High Titer | Lenti-p53 siRNA | High Titer Recombinant p53 siRNA Lentivirus (109 IU/ml) | 5 x 20 μl | LV619 |
| Lenti-Rb siRNA Virus | Lenti-Rb siRNA | Recombinant Rb siRNA Lentivirus (106 IU/ml) | 10 ml | G223 |
| Lenti-Rb siRNA Virus, High Titer | Lenti-Rb siRNA | High Titer Recombinant Rb siRNA Lentivirus (109 IU/ml) | 5 x 20 μl | LV621 |
| Lenti-Bmi1 Virus, High Titer | pLenti-Bmi1 | High Titer Recombinant Bmi1 Lentivirus (109 IU/ml) | 5 x 20 μl | LV608 |
| Lenti-Bmi1 Virus | pLenti-Bmi1 | Recombinant Bmi1 Lentivirus (106IU/ml) | 10 ml | LV610 |
| Lenti-CDK4 Virus, High Titer | pLenti-CDK4 | High Titer Recombinant CDK4 Lentivirus (109 IU/ml) | 5 x 20 μl | LV609 |
| Lenti-CDK4 Virus | pLenti-CDK4 | Recombinant CDK4 Lentivirus (106IU/ml) | 10 ml | LV611 |
| E1A-E1B Lentivirus | pLenti-E1A-E1B | Recombinant Adenoviral E1A-E1B Lentivirus (10 6 IU/ml) | 10 ml | LV651 |
| E1A-E1B Lentivirus, High Titer | pLenti-E1A-E1B | High titer recombinant Adenoviral E1A-E1B Lentivirus (10 9 IU/ml) | 5 x 20 μl | LV652 |
| Lenti-Myc T58A Virus | pLenti-Myc T58A | Recombinant Myc T58A Lentivirus (106 IU/ml) | 10 ml | G217 |
| Lenti-Myc T58A Virus, High Titer | pLenti-Myc T58A | High Titer Recombinant Myc T58A Lentivirus (109 IU/ml) | 5 x 20 μl | LV618 |
| Lenti-Ras V12 Virus | pLenti-Ras V12 | Recombinant Ras V12 Lentivirus (106 IU/ml) | 10 ml | G221 |
| Lenti-Ras V12 Virus, High Titer | pLenti-Ras V12 | High Titer Recombinant Ras V12 Lentivirus (109 IU/ml) | 5 x 20 μl | LV620 |
HOX Gene Cell Immortalization Reagents 基因不朽化商品
| Product |
Vector Map (Click to View) |
Description | Quantity | Cat. No. |
| Lenti-HOXA10-C-term-HA Lentivirus | pLenti-HOXA10-C-term-HA | Recombinant HOXA10 Lentivirus with HA (106 IU/ml) | 10 ml | LV645 |
| Lenti-HOXA10-C-term-HA Lentivirus, High Titer | pLenti-HOXA10-C-term-HA | High titer recombinant HOXA10 Lentivirus with HA (109 IU/ml) | 5 x 20 μl | LV648 |
| Lenti-HOXA10-GFP Lentivirus | pLenti-HOXA10-GFP | Recombinant HOXA10 Lentivirus with GFP (106 IU/ml) | 10 ml | LV646 |
| Lenti-HOXA10-GFP Lentivirus, High Titer | pLenti-HOXA10-GFP | High titer recombinant HOXA10 Lentivirus with GFP (109 IU/ml) | 5 x 20 μl | LV649 |
| Lenti-HOXA10-RFP Lentivirus | pLenti-HOXA10-RFP | Recombinant HOXA10 Lentivirus with RFP (106 IU/ml) | 10 ml | LV647 |
| Lenti-HOXA10-RFP Lentivirus, High Titer | pLenti-HOXA10-RFP | High titer recombinant HOXA10 Lentivirus with RFP (109 IU/ml) | 5 x 20 μl | LV650 |
| Lenti-HOXA9-C-term-HA Lentivirus | pLenti-HOXA9-C-term-HA | Recombinant HOXA9 Lentivirus with HA(106 IU/ml) | 10 ml | LV639 |
| Lenti-HOXA9-C-term-HA Lentivirus, High Titer | pLenti-HOXA9-C-term-HA | High titer recombinant HOXA9 Lentivirus with HA (109 IU/ml) | 5 x 20 μl | LV642 |
| Lenti-HOXA9-GFP Lentivirus | pLenti-HOXA9-GFP | Recombinant HOXA9 Lentivirus with GFP (106 IU/ml) | 10 ml | LV640 |
| Lenti-HOXA9-GFP Lentivirus, High Titer | pLenti-HOXA9-GFP | High titer recombinant HOXA9 Lentivirus with GFP (109 IU/ml) | 5 x 20 μl | LV643 |
| Lenti-HOXA9-RFP Lentivirus | pLenti-HOXA9-RFP | Recombinant HOXA9 Lentivirus with RFP (106 IU/ml) | 10 ml | LV641 |
| Lenti-HOXA9-RFP Lentivirus, High Titer | pLenti-HOXA9-RFP | High titer recombinant HOXA9 Lentivirus with RFP (109 IU/ml) | 5 x 20 μl | LV644 |
| Lenti-HOXB8-C-term-HA Lentivirus | pLenti-HOXB8-C-term-HA | Recombinant HOXB8 Lentivirus with HA (106 IU/ml) | 10 ml | LV633 |
| Lenti-HOXB8-C-term-HA Lentivirus, High Titer | pLenti-HOXB8-C-term-HA | High titer recombinant HOXB8 Lentivirus with HA (109 IU/ml) | 5 x 20 μl | LV636 |
| Lenti-HOXB8-GFP Lentivirus | pLenti-HOXB8-GFP | Recombinant HOXB8 Lentivirus with GFP (106 IU/ml) | 10 ml | LV634 |
| Lenti-HOXB8-GFP Lentivirus, High Titer | pLenti-HOXB8-GFP | High titer recombinant HOXB8 Lentivirus with GFP (109 IU/ml) | 5 x 20 μl | LV637 |
| Lenti-HOXB8-RFP Lentivirus | pLenti-HOXB8-RFP | Recombinant HOXB8 Lentivirus with RFP (106 IU/ml) | 10 ml | LV635 |
| Lenti-HOXB8-RFP Lentivirus, High Titer | pLenti-HOXB8-RFP | High titer recombinant HOXB8 with RFP Lentivirus (109 IU/ml) | 5 x 20 μl | LV638 |