What disease does lentivirus cause?
Lentivirus is a genus of retroviruses that cause chronic and deadly diseases characterized by long incubation periods, in humans and other mammalian species. The genus includes the human immunodeficiency virus (HIV), which causes AIDS.
What cells do lentivirus infect?
HIV-1 primarily infects cells that express the CD4 molecule, particularly CD4+ T lymphocytes. Other target cells include monocytes, macrophages, dendritic cells, astrocytes and microglia.
How does lentiviral infection work?
Lentiviruses (a genus of retrovirus) express reverse transcriptase, which converts the viral RNA to double stranded DNA, and integrase, which inserts this viral DNA into the host DNA. Once the viral DNA is integrated into the host DNA, it divides along with host cell and none are the wiser.
Can lentivirus infect mouse cells?
Yes, you can use lentivirus to stably transduce rodent primary cells. Although lentiviral vectors do transduce non-dividing cells, the efficiency is not always as good as with dividing cells. High multiplicity of infections (MOI) may lead to multiple integrations and very high levels of gene expression.
How is lentivirus transmitted?
Transmitted from person to person through direct exposure to infected body fluids (blood, semen) sexual contact, sharing unclean needles etc.; transplacental transfer can occur.
What is lentiviral infection?
Lentivirus infections are characterized by a prolonged incubation period between initial infection to occurrence of disease (from the Latin “lenti,” meaning “slow”).
Can lentivirus infect humans?
In addition, as LVVs are often designed to infect a broader range of human cells than HIV [eg, replacing the HIV envelope with vesicular stomatitis virus envelope glycoprotein (VSV-G)], the entire mucosal membrane of the tracheobronchial region can potentially be infected.
What are the two main functions of lentivirus recombination?
The Lentivirus – The Recombinant Lentivirus System
|LTRs||Contain sequences required for viral gene expression, reverse transcription, and integration|
|ψ||Required for packaging of the genomic transfer RNA|
|RRE||Rev response element. Required for processing and transport of viral RNAs|
How do you do lentivirus transduction?
- Thaw the lentivirus on ice. Mix 8 µl Polybrene (1 mg/ml aliquot) with 957 µl culture.
- The next day, exchange Lentivirus/Polybrene mixture by fresh culture medium. Incubate cells at standard cell culture conditions.
- concentrations range from 0.1-10 μg/ml. Replace the culture medium 48-72 hours.
How can lentiviral transduction be improved?
10 best practices for a good transduction efficiency
- 1- Know your cells.
- 2- Check the quality of your lentiviral vector batch.
- 3- Optimize the transgene expression cassette.
- 4- Make sure your titer is in TU/ml.
- 5- Follow storage and thawing requirements.
- 6- Monitor cells sensitivity towards transduction enhancers.
How do you deal with lentivirus?
Physical Safety Precautions for handling Lentivirus
- Open virus containing microtubes in the biosafety cabinet.
- Best to avoid using sharps or implement additional care to avoid autoinoculation.
- Precaution should be taken when the gene to be introduced is oncogenic or otherwise dangerous to humans.
Are lentiviral vectors infectious?
Lentiviruses and lentiviral vectors are transmissible through injection, ingestion, exposure to broken skin or contact with mucous membranes of the eyes, nose and mouth. Lentiviral vector systems are based on the genome of HIV, but many are pseudotyped to have a broad range of cell tropism for infection.
What kind of host does a lentivirus infect?
Lentivirus (lente-, latin for “slow”) is a group of retroviruses characterized for a long incubation period. They are classified into five serogroups according to the vertebrate hosts they infect: bovine, equine, feline, ovine/caprine and primate. Some examples of lentiviruses are Human (HIV), Simian (SIV) and Feline (FIV) Immunodeficiency Viruses.
How big is a lentivirus for gene delivery?
HIV-derived vectors are highly efficient vehicles for in vivo gene delivery. Lentiviruses are approx. 120nm in diameter, enveloped, and contain a nucleocapsid containing two copies of single-stranded positive-sense RNA.
Is it safe to use a third generation lentiviral vector?
Cis and trans -acting factors of the lentivirus are on separate plasmids depending on the Viral Vector generation. Third generation systems are currently the safest to use because the virus production is split across four plasmids. Packaging cells: 293T cells Lentiviral vectors infect dividing and non-dividing cells.
What’s the best way to get rid of lentivirus?
10% freshly prepared bleach solution or 70% ethanol. Heat at 56˚C for 30+ minutes. 90-99% reduction in several hours. At minimum, personnel are required to don gloves, closed toed shoes, lab coat, and appropriate face and eye protection prior to working with Lentivirus. Additional PPE may be required depending on lab specific SOPs.