ORFPOOLWG
MISSION® TRC3 Human Whole Genome Lentiviral ORF Pool
About This Item
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packaging
pkg of 8x25 μL (vials)
Quality Level
concentration
5x108 VP/ml (via p24 assay)
technique(s)
capture ELISA: 5 × 108 TU/mL using p24
shipped in
dry ice
storage temp.
−70°C
Related Categories
General description
This ready-to-use MISSION TRC3 Human LentiORF pooled library allow for stable integration, enrichment of cells, and long-term gene expression in difficult-to-transfect cell lines utilizing our best-in-class lentiviral manufacturing. This collection provides researchers with the unique tools to gain insights into gene function through modulating gene and protein expression.
The classical genetic approach for exploring biological pathways typically begins by identifying mutations that cause a phenotype of interest. Overexpression or misexpression of a wild-type gene product, however, can also cause mutant phenotypes, providing geneticists with an alternative yet powerful tool to identify pathway components that might remain undetected using traditional loss-of-function analysis. The whole genome lentiviral ORF pool fills this gap and provides researchers with a powerful research tool for carrying out crucial biological experiments.
Whether looking to perform gene overexpression studies or validate gene knockdown/knockout results from an RNAi or CRISPR experiment, LentiORFs are the ideal shortcut to protein expression and tool for gene analysis. Researchers also need to validate “hits” from a screen to confirm to eliminate false positives. This means confirming whether the observed phenotype is due the target gene of interest. Re-expressing the gene of interest via lentiORFs is the most accepted way to validate a gene target.
MISSION TRC3 Human LentiORF pooled whole genome library is provided in 8×25 μl aliquots at a minimum titer of 5×108 TU/ml (measured by a p24 assay).
Application
Features and Benefits
- Efficiently screen the whole human genome (14,000+ genes) at the bench-top without robotics or specialized equipment
- Pre-cloned, sequence confirmed, expression-ready clones
- Ideal Shortcut to Protein Expression: Use Pooled Lentiviral ORFs to express protein-coding genes to assess their function
- Simple Deconvolution: The puromycin library is barcoded, consisting of a 24 nucleotide sequence located downstream of the V5 tag and translation stop. Each clone contains a unique barcode sequence that can be easily identified, allowing for simple deconvolution of pooled screening experiments
- EF1a promoter drives expression of the ORF construct, offering an alternative to the CMV promoter which may not be appropriate in certain cell types
- Puromycin resistance allows for double selection experiments in cells containing Neomycin or Blasticidin resistance
- Enriched population of cells: Use puromycin selection after transduction, for creation of a long term-stable cell line
- V5 epitope tag facilities pull-down and co-immunoprecipitation experiments to determine protein-protein interactions or label and visualize ORFs via immunofluorescence
- Positive and negative selection controls (BFP and GFP) are provided, which allow researchers to confidently gauge the success of their pooled screening experiments
- Lentiviral ORFs can infect a broad variety of mammalian cells (including difficult to transfect or primary cell lines) via transduction to facilitate gene overexpression for screening applications
- Rapid and affordable screening with easy-hit identification via Sigma-Aldrich′s deconvolution service
- Save time: Skip PCR, cloning, and DNA sequence verification steps
Preparation Note
Other Notes
Legal Information
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
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Articles
Our lentiviral vector systems are developed with enhanced safety features. Numerous precautions are in place in the design of our lentiviruses to prevent replication. Good handling practices are a must.
Successful targeting relies on optimizing key sensitive steps in the process, including lentiviral transduction. Below are some helpful handling and titration tips from our R&D lentiviral experts.
Protocols
You are not alone designing successful CRISPR, RNAi, and ORF experiments. Sigma-Aldrich was the first company to commercially offer lentivirus versions of targeted genome modification technologies and has the expertise and commitment to support new generations of scientists.
FACS (Fluorescence-Activated Cell Sorting) provides a method for sorting a mixed population of cells into two or more groups, one cell at a time, based on the specific light scattering and fluorescence of each cell. This method provides fast, objective, and quantitative recording of fluorescent signals from individual cells.
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
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