Altogen Labs RNAi Services
Altogen Labs http://altogenlabs.com offers a multitude of RNAi (RNA interference) services http://altogenlabs.com/rna-interferen.... Our services include in vivo siRNA delivery and tissue targeting.
-In Vivo siRNA Delivery and Tissue Targeting (In Vivo RNAi)
Two basic methods are adapted for RNAi use in vivo: administration of siRNA or miRNA and administration of plasmid DNA and viral vectors, which in vivo express short hairpin RNAs (shRNAs) that are subsequently processed into active siRNA.
One of the key challenges faced when using RNAi technology is the successful delivery of a stable and functional siRNA or microRNA molecule to the target tissue. Altogen Labs has developed methods to prevent siRNA degradation in serum (including both chemical modification and encapsulation of siRNA), increasing delivery efficiency to the target organ, and producing strong tissue-targeted functional effects.
Chemical modifications of siRNA or miRNA are performed to increase their stability in vivo. These chemical modifications sustain the potency, efficacy and specificity of the siRNA molecules and increase their overall stability in the dynamic environment of a living organism. Standard modifications include 2’MOE, FANA, 2’-Fluoro, LNA, and encapsulation into liposomes.
-siRNA Transfection and RNAi Cell-based Library Screening
In the pursuit of understanding gene functionality, RNAi library screening has become a valuable research tool used to study gene function and associated signaling pathways. RNAi screening has allowed researchers to study the functionality of genes in a cell, both individually and in combinations of up to thousands of genes at a time, by temporarily silencing their expression.
Altogen Labs offers siRNA and miRNA library screening, primary (high-throughput) and secondary cell-based assay development, and validation. At Altogen Labs we have developed and validated highly efficient and optimized delivery systems for over 80 cell lines and over 20 primary cell cultures to provide high throughput shRNA, siRNA and miRNA library screening.
Custom RNAi services include: siRNA Transfection, RNAi cell-based library screening, siRNA/shRNA design, synthesis, in vitro validation, qRT-PCR test of target mRNA knockdown, development of RNAi cell lines, mRNA reduction in tissues, siRNA targeting, chemical modification, cell transfection optimization, siRNA and microRNA genome-wide library screening, RNAi assay development. Altogen Labs offers transient and stable RNAi transfection services based on the client’s individual needs.
-Generation of Tet-inducible RNAi Gene Knockdown Cell Lines
Altogen Labs offers the development of inducible RNAi stable cell lines expressing shRNA constructs that target genes of interest. The generation of tetracycline-inducible RNAi gene knockdown cell lines allows researchers to tightly control short hairpin RNA (shRNA) expression.
Altogen Labs scientists carry years of extensive cell culture experience and expertise, including complete RNAi services and generation of inducible cell lines (the client is only required to provide the gene ID of the target).
Standard services include transfection of plasmid DNA (at least 20 µg) into a cell line of interest, drug selection of clonal cells, generation of stable cells, validation of functional shRNA construct expression, and RNAi gene silencing by real-time qRT-PCR (reduction of mRNA expression) and/or Western blot (reduction of protein expression).
Final cell lines are delivered to the client as cryovials on dry ice.
-RNAi Screen Pathway Analysis: siRNA and microRNA
RNA interference (RNAi) is a powerful experimental method for specific silencing of genes, and is now an important technology for the elucidation of gene function, as well as gene involvement in specific biological processes.
RNAi screening technology (high-throughput siRNA and microRNA library screens) accelerates the identification of novel genes and their functions, and accordingly, the discovery of new gene signaling pathways.
Genome-wide library screening produces a comprehensive set of genes involved in cell signaling pathways, and enables secondary screens for more detailed pathway analysis and characterization.
With over 30,000 genes in the human genome, simultaneous RNAi library screening of several thousand genes requires robust statistical pipeline and pathway analysis.
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