What is virus pseudotyping?

Pseudotyping is the process of producing viruses or viral vectors in combination with foreign viral envelope proteins. The result is a pseudotyped virus particle, also called a pseudovirus.

Can you vortex lentivirus?

For each dilution, dilute the lentiviral stock into complete culture medium to a final volume of 1 ml. DO NOT vortex.

Why are lentiviruses used?

Lentiviral vectors have become particularly attractive for clinical applications due to their ability to more efficiently transduce non-proliferating or slowly proliferating cells, such as CD34 + stem cells.

How do you make lentivirus?

Procedure

  1. Seed 293T packaging cells at 3.8×106 cells per plate in DMEM complete in 10 cm tissue culture plates.
  2. Incubate the cells at 37 ℃, 5% CO2 for ~20 hours.
  3. Gently aspirate media, add 10 mL fresh DMEM complete containing 25 μM cloroquine diphosphate and incubate ~5 hours.

Can humans get vesicular stomatitis?

Humans can contract vesicular stomatitis by coming into contact with lesions, saliva, or nasal secretions from infected animals. In people, the disease causes an acute influenza- like illness with symptoms such as fever, muscle aches, headache, and malaise.

What kind of virus is VSV?

Vesicular stomatitis virus (VSV) is an enveloped, negative-sense RNA virus that infects a wide variety of mammalian and insect cells. Infections in humans are asymptomatic or result in a mild febrile illness. This virus is also exquisitely sensitive to interferons (IFN).

Can I freeze lentivirus?

Storage of Lentivirus Virus can be stored at 4°C for a short time (less than a week) before using after reception. Since Lentiviruses are sensitive to freeze-thawing and the titer drops with repeated freeze-thawing, aliquot viral stock should be stored at – 80°C freezer immediately upon arrival for long-term usage.

How can I improve my lentivirus titer?

found that in some cases, titers can drop 10-fold for every 2kb increase in insert size. To overcome this, lentivirus can be concentrated by a variety of methods such as ultracentrifugation, centrifugation on a sucrose gradient, spin columns, and polyethylene glycol (PEG) precipitation.

How do lentiviruses 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 humans?

Acute infection with human lentiviruses can appear as non-specific “flu-like” and “mononucleosislike” symptoms, including myalgia, arthralgia, diarrhea, nausea, vomiting, headache, hepatosplenomegaly, weight loss and neurological symptoms.

What is the difference between lentivirus and retrovirus?

Lentiviruses are a subtype of retrovirus. The main difference between lentiviruses and standard retroviruses from an experimental standpoint is lentiviruses are capable of infecting non-dividing and actively dividing cell types, whereas standard retroviruses can only infect mitotically active cell types.

What is Delta-G-VSV pseudotyping?

This Delta-G-VSV Pseudotyping System has proven useful for identifying cellular receptors for viruses, screening for entry inhibitors, and evaluating neutralizing antibody responses following vaccination. Originally described in a 2010 methods article ( PMID: 20709108 ), the system is now used as a virus model system by researchers worldwide.

What is PLP/VSVG plasmid?

Plasmid: pLP/VSVG. Lentiviral packaging plasmid for expression of the vesicular stomatitis virus G glycoprotein.

Can VSV-G envelope glycoprotein be used for pseudotyping of lentiviral vectors?

While the envelope glycoprotein of vesicular stomatitis virus (VSV-G) is widely used for pseudotyping of lentiviral vectors, sub-optimal gene transfer into certain cell types and its sensitivity to inactivation by human complement hinders its broader applications.

Why pseudotyping viral systems?

Therefore, pseudotyping viral systems have been widely employed to study highly infectious and pathogenic viruses such as Ebola virus, Middle Eastern Respiratory Syndrome (MERS) virus, or SARS viruses (12–14).