The Basics of the Recombinant Lentivirus System

How do recombinant lentivirus systems work? Lentiviruses are members of the Retroviridae family of viruses, with HIV-1 being the most widely studied. Lentiviral vectors are able to deliver genes into a broad range of mammalian cell lines and tissue types and to infect both dividing and non-dividing cells. Recombinant lentivirus systems and lentiviral transfection/transduction are powerful tools and methods for gene delivery and gene therapy. Because they can stably integrate transgenes into the host genome, lentiviruses are unique tools for generating stable cell lines for long term expression. In this video we will introduce you to the basic structure and genome map of the HIV Lentivirus and explain the differences and advantages of using first, second, and third generation recombinant lentivirus packaging systems. In the recombinant system, the HIV lentiviral genome is split into multiple plasmids (e.g. lentiviral transfer plasmids, packaging plasmids and envelop plasmids) that are delivered separately to ensure the introduced lentivirus cannot replicate in host cells. Thus, though natural lentiviruses such as HIV, SIV, FIV, EIAV, and Visna can cause chronic and even fatal diseases, scientists have managed to reduce the biosafety risks to a minimal level and convert this group of viruses into a tool for research and treatment of diseases such as cancers.

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[WulfLOL]

Very easy to understand and elegant way to present the different generations! ♥

[jliu]

Hi,
4:28 "self-inactivation system is created as the deletion is transferred to the 5' LTR after reverse transcription and integration"
Could you give more detail regarding how the deletion can be transferred to 5' LTR?

[samueleffah]

Hello. Would like to confirm if these recombinant lentivirus can infect an already infected CD4 Tcell especially those with reduced CD4 expression due to HIV accessory proteins.

[rbkuwar]

I am trying to clone transcription factors such as sox2, klf4 and oct 4 and c-Myc using lentivirus and retrovirus in the fibroblast culture. I am very new in this field. can you guide me from where I should start it? I am naive in the cloning work....

[shadowx]

Do lentivectors used in production of t-cell therapy cause false positive results in HIV testing?

[westfield]

Very interesting video. Now 4 years after this video. Have there been breakthroughs? Just hoping

[fireteamomega]

Impressive so its viral replication is controlled by removing rna transcription factors and removing the dimer making further synthesis impossible. Then you're adding a broad range receptor surface for enhanced binding. But how is exact delivery determined then ? Wouldn't there be a possibility of nullifying or cross linking proteins that have ambiguity in binding receptor surfaces ? I just imagined the capsid surface would have a designer site specific receptor like one from an antigen or antibody.

[quangvinhbui]

Hello, Can you answer this question that if non-engineered lentivirus don't have psi gene for packing then what gene they use to do that ?

[mindofzyzz]

How does lenti compare to AAV for crispr knockout in human?

[basilbaby]

Could you differentiate between the tranfer plasmid with and the envelope plasmid?

For example, if I want spike proteins on the pseudoviruses, to which plasmid( transfer plasmid or envelope plasmid ) should I clone the sequence for spike protein?

[rosreydy]

I wonder that, why 293 T cells were used in Lentivirus Production?

[linalinchen]

Hello,
what's with inserting more than 1 transgene? What do I have to consider about? And how many transgenes will have the "offsprings"?

[shayomiumanga]

is this process applicable for lenivirus simian virus also?

[suneethaindupalli]

Hi I tried with abm lenti vector with third generation packaging mix but GOI was not packaging into viral particles

[cacevedo]

Very enlightenment video. Many thanks!

[ousmanbajinka]

I need to develop pseudovirus for RSV; Any technical assistantance?

[JC-lypz]

Will Lentivirus spread to infect more cells? How effective is it to get to target tissue to have an effect with a single injection? Is does it need multiple injections?

[soulsymphony]

why do I keep hearing losing my religion in this video, haha anyways I have to thank you it's very elaborated