Available services performed by the experienced personnel at Altogen Labs include:

• Clonal cell line development
• Cell engineering
• Stable transfection
• Cloning of construct, subcloning, amplification of plasmid DNA and purification
• Identifying desired trait expression at varying levels in clones, isolation of preferred colonies

A typical project to produce cells that overexpress a protein (such as this study) may include:

• transfection of plasmid DNA,
• selection of clonal cells via reporter gene,
• colony pick up,
• stable cell line generation,
• expression and functional screening (a minimum of ten passages), and
• final validation of gene construct expression by qRT-PCR, Western blot analysis, or other technique

STABLE CELL LINES FOR RNAI RESEARCH AND DEVELOPMENT

Biomedical researchers and labs utilizing RNA interference technology often rely on stable cell lines for shRNA expression, shRNA vectors for long-term gene silencing. RNAi or RNA interference, is a process that engages gene expression, reducing the expression or formation of the short hairpin RNA or shRNA in a specific sequence within a cellular structure. Transient RNAi is known to utilize siRNA, while induction of stable RNAi require generation of targeted shRNA, cloning into expression vector, and stable expression in host cell line. Plasmid DNA construct stable transfection that contains shRNA construct enables researchers to manipulate gene expression of target gene and study it’s function in disease process or cell signaling, providing an important research tool utilized today in a number of therapeutic treatments, research and methodologies. However, the process of producing a stable RNAi cell line is time consuming, laborious, and often requires cell culture skills and extensive experience in the development of stable cell lines, although development of stable expression lines extremely important in extended research scenarios that often involve biological studies regarding the impact of protein and gene expression levels.

A study to produce an RNAi stable cell line (such as this project) may include:

• creation of plasmids encoding antibiotic-resistant genes and shRNA constructs that target the genes of interest,
• final plasmid DNA amplification,
• maxi-prep purification,
• transfection of DNA plasmid into the chosen cell line,
• drug selection,
• stable cell generation,
• final validation of gene construct expression by qRT-PCR, Western blot analysis, or other technique

The above are only suggested protocol steps; the experienced scientists at Altogen Labs work individually with each client to determine the best experiments for their needs.

REQUEST A FORMAL SERVICE QUOTE:

• Development of Stably Expressing Cell Line
• Stable RNAi Cell Line Development: shRNA Gene Knockdown
• Protein overexpression: Mammalian stable cell line development
• Generation of Stable Cell Line in 28 Days

BIOLOGICAL RESEARCH SERVICES | SCIENCE OF STABLE CELL LINE GENERATION | IN THE CLINIC AND LABORATORY | IN-HOUSE QUALITY CONTROL | COMPREHENSIVE CELL LINE DEVELOPMENT