Can we co-transduce one cell with two lentivirus of VSV-g

Virology Retrovirus (HIV)
1

Hello,

I heard VSV-g envelop can make multi-infection on single cell. But, I want to know how much cells can be co-transduced in especially primary cultured neurons.

I think that my successful experiments depend on the efficiency.

Please help me.

2

Barry

You’re correct that the Vesicular Stomatitis Virus glycoprotein (VSV-G) envelope enables broad tropism and efficient transduction of various cell types, including primary neurons. However, when assessing co-transduction efficiency (i.e., how many viral particles can infect a single neuron), several factors influence the outcome:

Factors Affecting Co-Transduction in Primary Neurons

  1. Multiplicity of Infection (MOI) – Higher MOI increases the likelihood of multiple viral particles entering a single cell.
  2. Neuronal Maturity – Younger neurons (DIV 3–7) may have different viral uptake capabilities than mature ones (DIV 14+).
  3. Lentivirus vs. AAV Systems – VSV-G pseudotyped lentiviruses can efficiently transduce neurons, but they do not replicate, limiting reinfection after integration. AAV vectors can be used for co-transduction but have different receptor requirements.
  4. Competition Among Viruses – If multiple VSV-G pseudotyped lentiviruses are used simultaneously, they may compete for entry receptors, reducing co-transduction efficiency.
  5. Neuronal Viability – Too high an MOI can induce cytotoxicity, especially in sensitive primary neurons.

Reported Co-Transduction Efficiency

  • In non-neuronal cells, studies show that a single cell can be infected by multiple VSV-G pseudotyped viruses when MOI is sufficiently high.
  • In primary neurons, co-transduction rates depend on MOI but are typically 30-80% efficient when two or more viruses are used at MOI ≥5.
  • Some studies using fluorescent protein markers (GFP, RFP, CFP) report 50-60% of neurons co-expressing two constructs when MOI is optimized.

Optimizing Co-Transduction in Primary Neurons

  • Use an MOI between 5 and 20, depending on viral titer and toxicity.
  • Transduce at DIV 5-7, when neurons are more permissive to infection.
  • Enhance viral uptake with polybrene (non-neuronal) or protamine sulfate (neuronal cultures).
  • Use different fluorescent markers to quantify co-transduction efficiency.
2 Likes