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A number of studies have demonstrated the successful application of RNAi in primary cell cultures and non-neuronal cell lines; however the use of gene silencing in neuronal cell types has been notoriously difficult because of several technical and biological limitations. Neurons have been considered the most resistant to RNAi. Methods to consider when performing nucleic acid delivery experiments with neuronal cultures are: electroporation, microinjection, viral-based methods, and certain chemical transfection reagents. Although physical delivery methods appear to be efficient for siRNA-induced studies where transient modification of gene expression is sufficient, these methods do not appear to be adequate for behavioral studies where long term alteration of gene expression may be necessary. The viral-based siRNA expression systems appear to be most applicable for both Central Nervous System (CNS) gene therapy and basic neurobiological studies, while adeno-associated virus (AAV) delivery systems enable long-term, stable gene expression with relatively little cytotoxicity. Lentiviral vectors have been reportedly used successfully for the delivery of short hairpin RNA (shRNA), a precursor of siRNA, into primary neurons to induce RNAi. The advantage of lentiviral system is the ability to transfect non-dividing cells.
Visit Altogen Biosystems Transfection Resource: RNAi applications (siRNA transfection, microRNA transfection, plasmid DNA transfection), stable and transient DNA transfection, delivery methods, electroporation, microinjection, dendrimer-based, non-viral and virus-mediated gene delivery, liposome and non-liposomal reagents. Transfection of cancer cell lines vs primary cultures. RNA Interference as a research tool for gene expression studies, gene silencing in neuronal cells, RNAi therapeutics and In Vivo Transfection Reagents from Altogen Biosystems.
Altogen Biosystems is a life sciences company dedicated to the development, marketing and manufacture of cell type specific transfection reagents. Efficient delivery of DNA and RNA molecules (siRNA, plasmid DNA, mRNA, microRNA) enabled by advanced formulation of reagents and peculiar design of transfection protocols. Altogen Biosystems offers a complete transfection system for a broad range of cell lines and primary cells. Reagents are functionally tested to be reproducible, serum compatible, induce low toxicity, and can be used for co-transfection experiments, and high throughput applications.
siRNA Transfection || Transfection Methods || Stable Transfection || RNAi || siRNA Library Screening || RNAi Therapeutics || Cell Transfection || In Vivo Transfection Reagents || Cell Line Specific Transfection Reagents
In Vivo Transfection Reagents || Fibroblast Cells Transfection Reagent || Astrocyte Cells Transfection Reagent || HEK-293 Transfection Reagent || MEF Transfection Reagent || HepG2 Transfection Reagent || CHO Reagent || MDA-MB Transfection Reagent || Cos7 Transfection Reagent || A549 Transfection Reagent || NIH3T3 Reagent || HUVEC Transfection Reagent || LNCaP Transfection Reagent || MCF-7 Reagent || PC-12 Reagent
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.
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