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Frequently Asked Questions: Custom Lentivirus Service

email info@creative-biogene.com Tel: 1-631-626-9181 Fax: 1-631-614-7828

1.Do I need to modify my lab for work with lentivirus?
2.What is the difference between 2nd generation and 3rd generation lentiviral systems?
3.How can lentiviral vectors be used to make stable cell lines?
4.What are the advantages of using lentivirus to generate stable cell line?
5.Is it feasible to express cDNA from a lentiviral transfer vector normally used for shRNA expression?
6.Where does the genetic materials insert into the genome when delivered with lentivirus?


1.Do I need to modify my lab for work with lentivirus?
According to the NIH Office of Biosafety, the NIH recommends preparing your lab for BSL-2 standards when using lentivirus.

2.What is the difference between 2nd generation and 3rd generation lentiviral systems?
Briefly, 2nd generation lentiviral systems use more HIV proteins (on fewer plasmids) in order to produce functional lentiviral particles than 3rd generation systems. Third generation lentiviral systems are considered safer than second generation systems, but may be more difficult to use because they require transfection with four separate vectors in order to create functional lentiviral particles. A 3rd generation transfer vector can be used with a 2nd generation packaging system, but a 2nd generation transfer vector cannot be used with a 3rd generation packaging system.

3.How can lentiviral vectors be used to make stable cell lines?
Many lentiviral vectors insert some selectable markers, such as the puromycin resistance gene, conferring resistance to antibiotics. If these antibiotics are added to the growth medium, they kill off any cells that have not incorporated the vector and those cells that survive can be expanded to create stable cell lines. Apart from that kind of lentiviral vectors, some lentiviral vectors have some other makers such as GFP. Cells which have incorporated the vector can be separated using FACS.

4.What are the advantages of using lentivirus to generate stable cell line?
Lentivirus has a much higher positive clone rate with the selectable marker. Using lentivirus to generate stable cell line can save much more time, cost and labor than transfection based stable cell line generation.

5.Is it feasible to express cDNA from a lentiviral transfer vector normally used for shRNA expression?
Yes. But the promoter within the transfer vector must be changed. Most shRNA expression requires use of a RNA pol III promoter and cDNA expression requires the use of a RNA pol II promoter.

6.Where does the genetic materials insert into the genome when delivered with lentivirus?
It is commonly believed that lentivirus integration is random in the genome. However, some articles suggest that lentivirus intergrates in active genes.

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45-1 Ramsey Road, Shirley, NY 11967, USA
Tel: 1-631-626-9181
Fax: 1-631-614-7828
Email: info@creative-biogene.com

Europe
Tel: 44-207-048-3343

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