The Lenti-X shRNA Expression System uses lentivirus to deliver short hairpin RNA (shRNA) expression constructs into cells for the purpose of suppressing specific genes via RNA interference (RNAi).

• The pLVX-shRNA2 Vector (Cat. No. 632179; Figure 1), which is available separately, provides shRNA and fluorescent ZsGreen1 coexpression to allow easy identification and sorting of transduced cells

Lenti-X HT Packaging System is Included

The system includes our Lenti-X HT Packaging System, which uses a novel, Tet-Off-based, 4th-generation packaging strategy to produce lentiviral supernatants with titers that are superior to most commercially available packaging systems. You can often transduce an entire dish of target cells with just 10¥ìl of unconcentrated viral supernatant. See our Lenti-X FAQ for more information.

High Titers Maximize Gene Knockdown

Given the varied efficacy of different shRNAs, high titers are essential to achieving the highest possible knockdown efficiency because they enable you to:

• Transduce 100% of your target cell population.
• Transduce every cell with sufficient copies for high-level shRNA expression.

Figure 2 demonstrates highly efficient luciferase knockdown in a population of HEK 293 cells transduced with a LVX-shRNA lentivirus.


Figure 1. Lenti-X pLVX-shRNA vectors for shRNA expression. The pLVX-shRNA1 vector offers puromycin selection capability and is included with the Lenti-X shRNA Expression System. pLVX-shRNA2 provides fluorescent protein coexpression (ZsGreen1) and is available separately (Cat. No. 632179).


Figure 2. A Lenti-X virus harboring a potent anti-luciferase shRNA expression construct effectively inhibited luciferase expression in HEK 293 cells. HEK 293 Tet-Off cells that stably express high levels of luciferase from pTRE-Luc were either untreated (MOI=0) or transduced with LVX-shRNA1 lentivirus (MOI=10) encoding an anti-luciferase shRNA. Cells were harvested 48 hr after transduction and then assayed for luciferase activity.

Applications

Produce recombinant lentivirus for delivering shRNA expression and gene-specific RNAi to target cells.

Components

• Lenti-X shRNA Expression System (Cat. No. 632177)
  pLVX-shRNA1 Vector (Cat. No. 632178; not sold separately)
  Lenti-X HTX Packaging Mix (Cat. No. 631248; not sold separately)
  Xfect (Cat. No. 631317)
  Tet System Approved FBS, US-Sourced (Cat. No. 631105)
  Lenti-X GoStix (Sample) (Cat. No. 631242; not sold separately)
• pLVX-shRNA2 (Cat. No. 632179)
  pLVX-shRNA2 Vector
  

Storage Conditions

Store all components at -20¡ÆC.