GeneNet™ siRNA Libraries - Profile 38,500 Genes at Once
Assay 200,000 siRNA sequences in a single screening
- Discover new therapeutic targets and diagnostic markers functionally involved in critical responses
- Decipher signaling pathways
- Take advantage of the only cost effective technology for genome-wide functional screening
- Efficently screen in almost any cell type or in vivo biological system
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With System Bioscience's (SBI's) ready-to-use prepackaged FIV-based GeneNet Lentiviral siRNA Libraries you can functionally profile thousands of genes by introducing and screening tens of thousands of siRNA sequences in a single experiment. Similar to the revolutionary genome-wide siRNA screening approaches recently described in Nature (March 25, 2004, p. 427 and p. 431), this technology enables high-throughput simultaneous screening for genes that functionally inhibit any selectable response or phenotype.
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 Ready-to-use, Prepackaged GeneNet Libraries
GeneNet libraries are constructed by ligating synthesized oligonucleotide siRNA templates with digested pFIV library vectors. Following construction, each library is packaged into highly transducible VSV-G pseudotyped lentiviral particles using HEK293 packaging cells that are co-transfected with the pFIV-PACK™ Lentiviral Packaging Plasmids. All GeneNet Libraries are provided in a prepackaged, transduction-ready, ready-to-use form.
Design and Selection of siRNA Sequences
GeneNet siRNA Libraries consist of many thousands of siRNA vector constructs --- 4-5 different constructs per target gene. In addition to designing the gene-specific siRNA template sequences to include characteristics like favorable GC content, minimial secondary structure, and no termination signals, GeneNet Libraries siRNA sequences were designed to hybridize to probes present on Affymetrix GeneChip™ Arrays. This design facilitates detection of the siRNA sequences selected in functional screening experiments.
Functional Profiling with GeneNet siRNA Libraries
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Transduce target cells with the packaged GeneNet Library. Transduction with high-efficiency, pantropic VSV-G pseudotyped lentiviral particles produce a target cell population where almost every cell expresses at least one siRNA.
If necessary, treat transduced cells to induce the desired phenotype (e.g., radiation or chemical exposure, apoptosis induction, differentiation).
Select or isolate the population cells with desired phenotype after treatment (e.g., growth selection, cell sorting).
Recover siRNA templates sequences from selected target cells using PCR vector primers to known vector sequences flanking the siRNA inserts.
Identify the specific species of siRNA recovered from the selected target cell population by sequencing or by hybridization to an Affymetrix GeneChip® Array that recognizes all the siRNA templates present in the library. experiments.
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Screening for Genes Involved in Radiation Resistance
Cells were transduced with a library of 7,500 siRNA sequences targeting 1,500 genes. The expressed siRNA sequences were rescued from irradiated cells (treated) and control cells, and the siRNA template sequences present in each population were compared by microarray hybridization. Red indicates sequences enriched 5 - fold and blue indicates those enriched 3 - fold. These enrichments indicate siRNA targeting genes that make the cells radiation sensitive.
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GeneNet™ siRNA Libraries
| GeneNet™ Library |
Vector |
# Target
Genes |
# Target
Transcripts |
# siRNAs |
Cat. # |
| Human 8.5K |
pFIV-H1-copGFP |
8,400 |
8,500 |
43,800 |
SI201A-1 |
| Human 8.5K |
pFIV-H1-Puro |
8,400 |
8,500 |
43,800 |
SI202A-1 |
| Human 50K |
pFIV-H1-Puro |
38,500 |
47,400 |
200,000 |
SI206A-1 |
| Mouse 40K |
pFIV-H1-Puro |
34,000 |
39,000 |
150,000 |
SI222A-1 |
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SBI
