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shRNA

LentiExpress™ Technology

 

LentiExpress Human Kinome RNAi Screens – Now 40% off using promo code SFL
The Fastest Route to Viral-Based shRNA Gene Silencing.
Add your cells and go!

Would you like to conduct rapid, cost-effective lentiviral-based shRNA screens in primary or difficult to transfect cells? Are you interested in longer term or stable gene silencing while eliminating intensive reagent preparation of viral vectors and screen optimization parameters? Sigma Life Science provides an innovative solution with our LentiExpress technology.

LentiExpress screens consist of pre-arrayed lentiviral particles harboring shRNAs of The RNAi Consortium collection. We offer this premier ready-to-use product format for the Human Kinase shRNA collection, so you can simply add your cells and proceed directly to loss-of-function screens!

 

  • Economical – Single use format
  • Simple, Streamlined Procedure – No viral particle preparation. Add your cells and go!
  • Save Time – Get your assay results 1 month earlier
  • Robust – Increased transduction efficiencies and no extra freeze-thaw cycles
  • Comprehensive – Average of ~6 constructs per target gene

Go directly to:
Overview of The LentiExpress Method
LentiExpress Human Kinase shRNA Set
LentiExpress Optimization Plate
Data Using LentiExpress Technology
Ordering Information

Collaborator provided protocols and data:
LentiExpress Screen

Overview of The LentiExpress Method

MISSION® LentiExpress Human Kinase shRNA Set – Now 40% off using promo code SFL

Protein kinases are among the largest and most well studied gene families. Protein phosphorylation plays an essential role in intercellular communication in eukaryotic organisms by mediating signal transduction during development, transcription, immune response, metabolism, apoptosis, and cell differentiation. Aberrant regulation of kinases plays a causal role in many diseases, and the study of these proteins and their functions will contribute to the discovery and development of new therapeutics.

The MISSION LentiExpress Human Kinase shRNA Set contains ~3,200 lentiviruses carrying shRNA sequences targeting ~501 human kinase genes allowing for quick, high throughput loss-of-function screens. Each gene is represented by several individual constructs, targeting different regions of the gene sequence. The particles are pre-diluted to approximately 5,000 viral particles per well in a single reaction volume of 30 µL. In addition to the virus generated from the Human Kinases (K), each plate includes negative controls to monitor transduction efficiency.

The LentiExpress Human Kinase shRNA Set includes:

  • shRNAs targeting ~501 human kinase genes
  • ~3,200 individual lentiviral shRNA clones
  • 41 ready-to-use Pre-arrayed 96-well plates
  • Up to 80 shRNA constructs per plate
  • Negative controls in outer columns

Representative Plate Layout image

MISSION LentiExpress Optimization Plate – Now 40% off using promo code SFL

Optimization of transduction conditions is a central component of successful delivery using the LentiExpress method. For maximal transduction and knockdown, it is important to determine the optimal ratio of virus to cells, or the MOI (multiplicity of infection), for each new cell type.

The LentiExpress Optimization Plate accelerates this process allowing rapid determination of the optimal cell density for transduction. The 96-well plate contains pre-arrayed aliquots of TurboGFP™ Control Transduction Particles, allowing the user to simply add the desired numbers of their cell type of interest. Efficiency of transduction is subsequently monitored by assessment of GFP fluorescence.
The LentiExpress Optimization Plate includes:

  • TurboGFP Control Particles for quick visual monitoring
  • Non-target shRNA Control Particles
  • Empty Vector Control Particles

Optimization Plate Layout image

LentiExpress Method for Screening Tumor Suppressor Gene Set

The MISSION TRC Human Tumor Suppressor Gene Family Set (Product No. SH0531) was used as a proof of concept for the LentiExpress format. Scientists at Sigma-Aldrich were successfully able to screen for genes that could modulate Paclitaxel sensitivity. The results of this study were published in Oncology Reports. Pre-arrayed lentiviral particles were used to transduce A549 cells in 96-well plates and knockdown the corresponding tumor suppressor gene expression. Cells were split into two samples. One sample set was treated with Paclitaxel and the second set received medium only. Results were normalized to pLKO.1-puro empty vector control virus (Product No. SHC001V). Outliers were observed that either conferred more resistance or sensitivity to Paclitaxel treatment.

Cell Growth Chart image

Gene Knockdown Chart image

Multiple clones for four genes confirmed the LentiExpress method. The clones conferred increased cell sensitivity to drug treatment. These genes have the potential to be prognostic markers in chemotherapeutic responses.

Ordering Information

 

Product No. Product Name
SHX001-1EA MISSION LentiExpress Human Kinases
SHX001-3EA MISSION LentiExpress Human Kinases (For triplicate experiments)
SHXC01-1EA MISSION LentiExpress Optimization Plate
SHXC01-3EA MISSION LentiExpress Optimization Plate


LentiExpress product image 

The LentiExpress products are in stock and ready for shipment 24 hours after your order is placed. Please note that these products are shipped on dry-ice and require immediate storage at -70°C.


Contact Us

For questions about the library, pricing and quotes or other concerns, please e-mail us at: MISSIONRNAi@sial.com.

MISSION is a registered trademark of Sigma-Aldrich Co. LLC Label License.
LentiExpress is a trademark of Sigma-Aldrich.
TurboGFP is a trademark of Evrogen.

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