Stable RNAi Cell Line Development: shRNA Gene Knockdown

Stable RNAi Cell Line Generation: shRNA-induced stable gene knockdown

Gene silencing or gene knockdown, also known as RNA interference (RNAi), is a biological phenomenon in which gene expression is silenced due to the specific degradation of the corresponding messenger RNA (mRNA). This process occurs when double-stranded RNA (dsRNA), in combination with RNA-induced silencing complex (RISC), stimulates the sequence-specific reduction of protein production of a particular gene or genes. Gene knockdown is useful for many applications, including functional analysis of genes, gene target discovery, validation of gene targets, assay development, and screening of drug compounds.

Altogen Labs offers development of inducible RNAi stable cell lines expressing small hairpin RNA (shRNA) constructs that target genes of interest. These stable cell lines expressing shRNA constructs are important expression research tools for studying gene function in mammalian cells. However, generation of shRNA-expressing and tetracycline-inducible RNAi stable cancer cell lines can be complex due to the large number of required controls (including, for example, efficiency of gene silencing, controlling for off-target effects, and promoter compatibility).

Altogen Labs can generate stable knockdown (RNAi) cell lines based on individual client requirements and project specifications. Our scientists offer years of extensive cell culture experience and cell line development expertise.

Standard RNAi stable cell line development services include:

a) Cloning — 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, and

b) Stable cell line generation — transfection of DNA plasmid into the cell line of choice, drug selection, generation of stable cells, validation of shRNA construct expression, and RNAi gene silencing by real-time RT-PCR (mRNA expression) and/or Western blot (protein expression).

Altogen Labs’ life science research services include a wide range of pre-clinical studies, biology CRO services, xenograft and in vivo tox studies, custom liposome encapsulation, in vivo siRNA encapsulation and tissue-targeted delivery, as well as complete RNAi gene silencing services that include gene targeting, cloning of shRNA and microRNA into plasmid of choice, development of TET-inducible RNAi stable cell lines, siRNA synthesis and chemical modification, functional in vitro validation, and other gene silencing services.

Download Altogen Labs Stable Cell Line Development PowerPoint Presentation: [PPT]

At Altogen Labs, we recognize that providing fast delivery of products and services is critical to expediting research. We are committed to meeting our clients’ timely needs while maintaining high quality and integrity. All of our services undergo rigorous QC procedures.

Once we know the details of your project, we can provide you an immediate price quote (contact us via e-mail at info@altogenlabs.com or call Altogen Labs at 512-433-6177). Please note that experimental details will help us to provide an accurate quote and timeline estimate.

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Generation of Stable Cell Line in 28 Days

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