Stable RNAi Cell Line Generation: Stable Gene Knockdown
RNA interference (RNAi) is a cellular process in which gene expression is reduced in a sequence specific manner following the expression of short-hairpin RNA (shRNA) within the cell. Transfection of a plasmid DNA construct containing shRNA and an antibiotic resistant gene induces the knockdown gene expression of the gene of interest and enables selection and isolation of successfully transfected cells. Cells with a prolonged ability to proliferate in the presence of a selecting agent (antibiotic) are capable to initiate a stable RNAi gene knockdown cell line. This important research tool has been vital to the studying of mammalian gene function and gene targeting for potential therapeutic treatments.
Altogen Labs offers generation of stable RNAi cell lines expressing shRNAs targeting genes of interest. Such stable RNAi cell lines can be a powerful research tool, however, the generation of RNAi cell lines and primary cells can be very laborious, expensive, and time-consuming to produce. Development of stably expressing cell lines is important for long term research involving the investigation of the biological impact of gene and protein expression levels. The distinction of a stable cell line is that the exogenous genetic material (in this case shRNA) has been fully incorporated into the genome of the cell resulting in permanent expression of the vector (plasmid DNA construct).
Altogen Labs offers the generation of stable knockdown (RNAi) cell lines based on individual client requirements and project specifications. Validation of gene silencing is performed using real-time qRT-PCR (mRNA expression level) or Western blot (protein expression level). Company scientists bring years of extensive cell culture experience and cell line development expertise.
Standard service include:
a) Cloning and subcloning (vector construction)
• Plasmid DNAs encoding antibiotic-resistant genes
• Synthesis and cloning of shRNA constructs targeting genes of interest into plasmid DNAs
• Final plasmid DNA amplification, and Maxi-prep purification
b) Stable cell line development
• Transfection of plasmid DNA into the cell line of choice
• Drug selection
• Generation of stable cells
• Validation of shRNA construct expression
• Validation RNAi gene silencing by real-time qRT-PCR and/or Western blot
For an immediate price and timeline quote, please contact us at firstname.lastname@example.org or call Altogen Labs technical support at 512-433-6177. We will discuss your experimental details and any special needs, and provide an accurate quote and timeline estimate.
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 | Genome-wide siRNA and microRNA High-throughput Screens | Stable RNAi Cell Line Generation: Stable Gene Knockdown | siRNA Liposome Encapsulation Service | Transient (siRNA) and Stable (shRNA) Transfection Services