- Lentivirus나 MMLV retrovirus의 더 빠른 transduction 유도
- Stem cell과 같이 민감한 세포에 이상적
- Starter Kit은 magnetic separator를 포함한다
Lenti-X Accelerator는 lentivirus와 MSCV retrovirus를 포함한 MMLV retrovirus의 transduction을 촉진하도록 디자인된 magnetic bead 기반의 기술이다. Lenti-X Accelerator는 bead들 virus와 결합시키기 위해 20분동안 preincubation한 뒤, 단지 5분 동안만 세포를 바이러스 상층액(viral supernatant)에 노출시키기 때문에 매우 민감한 세포의 적용에 이상적이다. 반면 세포에 polybrene을 처리하면 하루밤동안 incubation을 해야한다.
Figure 1. Transduction with Lenti-X Accelerator. Placing virus-bound magnetic beads in a magnetic field greatly increases the localized virus concentration at the surface of the cell monolayer.
This reduces the transduction time to just 5 min after a 20 min preincubation to bind the beads to the virus-compared to transduction overnight with Polybrene. Accelerated transduction also limits exposure of your sensitive target cells to viral supernantant.
Figure 2. Lenti-X Accelerator provides high transduction efficiency in a 25 min protocol.
A ZsGreen1-expessing Lenti-X vector was used to transduce HT1080 cells. Lenti-X Accelerator beads (8 ㎕) were preincubated for 20 min at room temperature with 200 ㎕ (approx. 1x106 total IFU) of viral supernatant, and applied to HT1080 cells on a Magnetic Separator for 5 min. HT1080 cells were also transduced with the same amount of virus in the presence of 6 ㎍/ml Polybrene, for 5 min or overnight. After the cultures were grown for an additional 72 hr at 37℃, the number of transduced cells was determined by flow cytometry.
