Application

Functional Genomics/Overexpression Screening/Target Validation

General description

The MISSION TRC3 ORF GFP Lentivirus Control Transduction Particles contain an ORF insert expressing TurboGFP^™, and is useful as a positive (or negative control) in experiments using the MISSION TRC3 LentiORF Library Collection in arrayed or pooled format, including our latest Whole Genome LentiORF Pool. TurboGFP is an improved variant of the green fluorescent protein copGFP cloned from copepoda Pontellina plumata.The TurboGFP ORF control transduction particles are produced from the sequence-verified lentiviral plasmid, pLX307-puro-TurboGFP (ORFGFP). This positive control can be used for measuring transduction efficiency and optimizing ORF delivery in cell lines that do not survive any puromycin exposure. The GFP Control contains a gene encoding TurboGFP driven by the EF1a promoter. This control provides fast visual confirmation of successful transduction. Ampicillin and puromycin antibiotic resistance genes provide selection in bacterial or mammalian cells respectively. Unlike murine-based MMLV or MSCV retroviral systems, lentiviral-based particles permit efficient infection and integration of the construct into differentiated and non-dividing cells, such as neurons and dendritic cells, overcoming low transfection and integration difficulties when using these cell lines. Self-inactivating replication incompetent viral particles are produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids.In addition, the Control Transduction Particles are pseudotyped with an envelope G glycoprotein from Vesicular Stomatitis Virus (VSV-G), allowing transduction of a wide variety of mammalian cells. 200 µl of 106 TU/ml (via p24 titering assay) lentiviral particles are provided as frozen stock.When conducting experiments using MISSION TRC3 ORF clones, the proper controls should be a key element of your experimental design to allow for accurate interpretation of gene and protein expression results. The MISSION Control Transduction Particles are a critical positive control to monitor transduction efficiency.To see application data, protocols, vector maps, and more visit the TRC3 Information Page

Legal Information

MISSION is a registered trademark of Sigma-Aldrich Co. LLC

TurboGFP is a trademark of Evrogen Co.

Use of this product is subject to one or more license agreements. For details, please see the MISSION License Page

Preparation Note

Puro Kill Curve and Determining CFU (Colony Formation Unit) per mL. Prior to performing a library-scale screening, two preliminary experiments must be conducted. Visit the Lentiviralpool Information Page or refer to the technical bulletin.