RNAi Resource: siRNA Transfection

RNA Interference (RNAi) is utilized in many eukaryotic cells including those found in mammals, which enables protein complex called RISC associate with double-stranded RNA molecules and activate sequence-specific gene silencing (target mRNA degradation).

The chain of reactions involved in cells in this manner enables DNA to be transcripted into RNA, which is then translated into proteins. RNAi research is a valuable process in gene expression studies not only for the studies using cell cultures and living organisms (using in vivo siRNA transfection systems), but in synthetic production of the siRNA that may eventually be introduced into cells to induce gene suppression as disease-gene-targeted RNAi Therapeutics. A number of biology CRO companies are now providing RNAi Therapeutics development services.

siRNA molecules are not stable in animals (degrade in a matter of minutes) and therefore require siRNA encapsulation to be protected from degradation in vivo. Liposomes can be chemically modifoed to allow tissue targeting and delivery of functional siRNA in vivo (targeted siRNA delivery to animal cells and tissues).

RNAi has also been known as post-transcriptional gene silencing, and in 2006, two scientists by the name of Andrew Fire and Craig Mello earned the Nobel Prize in physiology and medicine for their work in RNAi studies.

Transfection of siRNA is used in biology laboratories to experimentally induce gene silencing (RNAi) in cultured cells in vitro and animals in vivo. Scientists perform gene expression studies, functional genomics, gene characterization, and discovery of signalling gene pathways by introduction of extracellular molecules, such as small interferring RNA (siRNA), microRNA (miRNA), plasmid DNA expressing shRNA, messenger RNA (mRNA).

Transient transfection term refer to introduction of siRNA (or miRNA) molecule inside the cell without affecting its genomic DNA (transient expression), while stable transfection happens at incorporation of plasmid DNA into genome of the host cell (stable expression). Altogen Labs is a Contract Research Company (CRO) offers pre-clinical research studies, GLP-compliant in vivo and in vitro research services, custom liposome encapsulation, cell banking, and provides development of stable cell line service in only 28 days.

There are various cell transfection methods, including chemical transfection, electroporation, sonoporation, magnetofection, and gene gun injection. Commonly used transfection kits and reagents are nanoparticle-, lipid-, liposome-, and polymer-based. Pre-optimizized transfection kits and products are manufactured by Altogen Biosystems.

Transfection Links:
Biology Research Resource: RNAi, Cell Transfection, Protocols, Methods
WikiPedia: What is transfection?
Biology Research Services: Stably-Expressing Cell Line Generation
In Vivo siRNA Transfection Kits and Reagents

·      IN VIVO TRANSFECTION KITS

·        IN-VIVO TRANSFECTION KITS:

·        Nanoparticle In Vivo Transfection Reagent

·        LIPID In Vivo Transfection Reagent

·        PEG-Liposome In Vivo Transfection Reagent

·        POLYMER In Vivo Transfection Reagent

·        Pancreas In Vivo Transfection Kit

·        Liver In Vivo Transfection Kit

·        Kidney In Vivo Transfection Kit

·      CELL LINE TRANSFECTION KITS

·        Products 293 – 786-O

·        Products A204 – Astrocyte

·        Products B16-F10 – BxPC-3

·        Products C2C12 – CT26.WT

·        Products D-407 – Fibroblast

·        Products H9c2 – HUVEC

·        Products IMR-90 – LS-174T

·        Products MCF-7 – MRC5

·        Products NCI-H1299 – NTERA-2

·        Products PANC-1 – RD

·        Products SK-BR3 – SW480

·        Products T98G – ZR-75-1

·      PRIMARY CELLS TRANSFECTION KITS

·        ASMC Transfection Kit (Aortic Smooth Muscle Cells)

·        Astrocyte Transfection Reagent

·        BJ Transfection Reagent (Skin Fibroblast Cells)

·        CFPEo (HTRpC) Transfection Kit (Trachea Epithelium)

·        Chromaffin Transfection Reagent (Chromaffin Cells)

·        DI-TNC1 Transfection Kit (Rat Brain Astrocytes)

·        Fibroblast Transfection Reagent (Fibroblast Cells)

·        HCAEC Transfection Kit (Coronary Artery Endothelial)

·        HCN-1A Transfection Reagent (Brain Neuronal Cells)

·        HMEC Transfection Kit (Mammary Epithelial Cells)

·        HUVEC Transfection Kit, Umbilical Vein Endothelial Cell

·        Keratinocyte Transfection Reagent (Keratinocytes)

·        MDCK Transfection Reagent (Kidney Cells, CCL-34)

·        MEF Transfection Reagent (Mouse Fibroblast Cells)

·        Neuro-2a Transfection Kit (Neuroblastoma, CCL-131)

·        SKNAS Transfection Kit (Neuroblastoma Cells)

·        SK-N-MC Transfection Kit (Neuroblastoma Cells)

·      ELECTROPORATION PRODUCTS

·        CCRF-CEM Electroporation Kit (CML Leukemia Cells)

·        Daudi Electroporation Kit (Burkitt’s Lymphoma Cells)

·        HL-60 Electroporation Kit (Plasmacytoma, CCL-240)

·        Jurkat Electroporation Kit (T-Cell Leukemia, TIB-152)

·        K562 Electroporation Kit (AML Leukemia, CCL-243)

·        KG-1 Electroporation Kit (Leukemia Cells, CCL-246)

·        Raji Electroporation Kit (Burkitts Lymphoma, CCL-86)

·        Ramos Electroporation Kit (Lymphoma Cells, CRL1596)

·        RAW 264.7 Electroporation Kit (Lymphoma, TIB-71)

·        RBL-2H3 Electroporation Kit (Basophilic Leukemia)

·      TRANSFECTION CONTROLS (GFP PLASMID, SIRNAS)

·        Apoptosis Inducing siRNA

·        Cell Cycle Arrest siRNA (5 nmol)

·        DNA Electroporation Buffer (30 ml)

·        GFP-expressing plasmid DNA (25 µg)

·        Non-silencing control siRNA

·        Protein Electroporation Buffer (30 ml)

·        siRNA Electroporation Buffer (30 ml)

·      TRANSFECTION EQUIPMENT AND SUPPLIES

·        Electroporation Cell Array System

·        NCI-60 Tumor Cell Line Transfection

·        Custom Targeted Transfection System