Generation of Tetracycline-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 regulate and control short hairpin RNA (shRNA) functionality. The regulation of the knockdown shRNA expression can be crucial when the functionality of the target gene is necessary for survival, metabolism, and other vital biological processes. The generation of inducible RNAi stable cancer cell lines can be very complex and laborious to produce with all the necessary controls involved (e.g., efficiency of gene silencing, control for off-target effects, and promoter compatibility).
Altogen Labs scientists bring 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 gene ID to target). Development of inducible shRNA-expressing stable cell lines involves the synthesis of multiple shRNA molecules, cloning into plasmid DNAs (encoding antibiotic-resistant gene for selection), in vitro testing of shRNA-expressing constructs for gene silencing efficiency (performed via transient transfection and qPCR-based analysis of target mRNA knockdown), final plasmid DNA amplification and stable cell line development (drug selection, clone isolation, stable cell line generation and characterization, 10 passages validation).
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 .
Altogen Labs provide CRO contract research services, including variety of preclinical studies, stable cell lines, cell banking services, and complete spectrum of RNAi gene silencing services: gene targeting, siRNA synthesis, chemical modification, functional in vitro validation, siRNA encapsulation, in vivo siRNA protection and tissue targeted delivery.
Please let us know the details of your project, so that we can provide you an accurate quote and timeline estimate (contact e-mail: firstname.lastname@example.org or call Altogen Labs technical support at 512-433-6177).
In Vivo siRNA Delivery and Tissue Targeting (In Vivo RNAi) | siRNA Transfection and RNAi Cell-based Library Screening | Generation of Tet-inducible RNAi Gene Knockdown Cell Lines | RNAi Screen Pathway Analysis: siRNA and microRNA | RNAi Manufacturing and CRO Services | PCR Cloning: DNA Vector Construction (shRNA) | Quantitation sh/siRNA-induced Knockdown by qRT-PCR and WB | Transient (siRNA) and Stable (shRNA) Transfection Services