Generation of Stably Expressing Cell Lines

Altogen Custom Services offer generation of stable cell lines by transforming your cell line to stably express vector or gene of interest. Plasmid DNA construct should encode at least one antibiotic resistance gene to enable drug selection. Transfected genetic material can be expressed in the target cells either transiently or permanently depending on the methods utilized and the experimental questions being investigated. Transient transfections are used most commonly to analyze the short term impact of altered gene or protein expression. Plasmid DNA (pDNA), messenger RNA (mRNA), short interfering RNA (siRNA), and microRNA (miRNA), are introduced and gene products are expressed in the target cells however the nucleic acids do not integrate into the host cell genome. Therefore, gene product expression is transient and typically results in high expression levels that persist for 24-72 hours when RNA is transfected, or 48-96 hours following DNA transfection. Conversely, in order to analyze the long term impact of altered gene or protein expression investigators typically utilize stable transfection protocols to develop stable cell lines. In a subpopulation of transfected cells, whether the desired effect is a stable or transient transfection, the transfected genetic material will integrate into the genome. In order to create stable cell lines, investigators will take advantage of this natural occurrence, and introduce the gene of interest along with a selectable marker. Therefore growth of transfected cells, in the presence of a selecting agent, will enable the subpopulation of cell where the exogenous genetic material has been incorporated into the genome to persist while the remaining cells undergo selection. Utilizing this method, investigators are able to develop cells that permanently express specific genes through their incorporation in the cellular genome.

For more information please contact us at corporate@altogen.com. Experimental details will help us to provide an accurate quote.

RNAi Services

RNA interference (RNAi) is a phenomenon by which the expression of double stranded RNA (dsRNA) specifically stimulates a cellular process that reduces gene expression in a sequence specific manner. Small synthetic RNA, termed small interfering RNAs (siRNA), which are typically 21-28 nucleotides in length, can induce RNAi and knockdown gene expression in mammalian cells without inducing an antiviral response. Gene silencing by RNA Interference (RNAi) is a powerful research tool for studying gene function in mammalian cells. RNAi is a biological phenomenon by which double stranded RNA (dsRNA) specifically reduces gene expression of its corresponding gene. Potent inhibition of specific gene expression is experimentally achieved by the transfection of small interfering RNA (siRNA), which binds RISC complex and cause degradation of target complementary mRNA molecules in the cell. Therefore, successful, potent RNAi experimentation is dependent upon the highly efficient delivery of the siRNA into cells by transfection of stable and functional siRNA molecules. Altogen Custom Services offer following RNAi Services:

• siRNA transfection optimization service
• Design and synthesis of siRNA targeting gene of interest
• Development of transfection reagents
• Gene of interest knockdown service

Please contact us at corporate@altogen.com for more information. Experimental details will help us to provide an accurate quote.

Custom Services

Custom laboratory research services include:

• Transfection services (transient and stable)
• Assay development and library screening
• Cloning and plasmid vector construction
• Generation of stable cell lines
• RNAi experiments (siRNA design, synthesis, validation)
• Gene targeting (mRNA knockdown, shRNA overexpression)
• Gene of interest knockdown service
• Custom services

For more information please contact us at corporate@altogen.com. Experimental details will help us to provide an accurate quote.

Transfection Resource

Visit Altogen's Transfection Resource: RNAi applications, stable and transient transfection, delivery methods, electroporation, nanoparticle, microinjection, dendrimer-based, non-viral and virus-mediated gene delivery, liposome and non-liposomal reagents, In Vivo Transfection Reagents from Altogen Biosystems.

Featured Products

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Altogen Research Services

Altogen Custom Services provide specialized biotechnology and pharmaceutical services, including - generation of stable cell lines, RNA Interference (RNAi) services, assay development, screening and transfection services. Generation of stably-expressing cancer cell lines and primary cells can be very expensive and time-consuming. Altogen Services offers generation of stable cell lines by transforming the cell line of choice to stably express gene of interest or shRNA-encoding construct. Cloning and shRNA expression services are available.