Accell GAPD Control siRNA

Validated Accell positive controls provide a reliable benchmark for RNA success.


Validated positive control siRNA targeting the GAPD housekeeping gene in human, mouse, or rat. Accell siRNA requires no transfection reagent or viral vector for delivery into difficult-to-transfect cells.

Accell GAPD Control siRNA is validated, highly reliable positive control for delivery and RNAi efficiency. Accell GAPD Control siRNA is modified with patent-pending Accell modification pattern to enable uptake by difficult-to-transfect cells. Researchers around the world are achieving targeted gene silencing in neuronal, immune, and primary cells that had previously been beyond the reach of conventional RNAi products due to toxicity caused by transfection reagents or undesirable viral responses.

GAPD (glyceraldehyde-3-phosphate dehydrogenase, also known as GAPDH) is a well-conserved enzyme in carbohydrate metabolism. This gene is abundantly expressed in most cells, and because it is non-essential, knockdown of the corresponding mRNA does not affect cell viability.

Highlights

  • Accell siRNA enters cells without the need for transfection reagents, virus (or viral vectors), or instruments
  • Novel siRNA modifications facilitate uptake, stability, specificity, and knockdown efficiency
  • Targets accession number NM_002046 (Human) or NM_008084 (Mouse) or XM_575242 (Rat)

Experimental considerations

  • Accell siRNA works at a higher concentration than conventional siRNA; recommended 1 µM working concentration
  • Delivery may be inhibited by the presence of BSA in serum. Optimization studies with serum-free media formulations (Accell Delivery Media) or < 2.5% serum in standard media is recommended
  • Full-serum media can be added back after 48 hours of incubation. Optimal mRNA silencing is typically achieved by 72 hours or up to 96 hours for protein knockdown
  
HazardousNo
Shipping ConditionAmbient
Stability at Recommended Storage ConditionsAt least 12 months
Storage Condition-20 C
Cell types demonstrating effective silencing with Accell siRNA

Cell types demonstrating effective silencing with Accell siRNA

Cell types demonstrating effective silencing with Accell siRNA

Internal validation and peer-reviewed publications report numerous successes with difficult-to-transfect cell types. See the References tab for a list of publications.


The Accell siRNA application protocol simplifies targeted gene knockdown

The Accell siRNA application protocol simplifies targeted gene knockdown

The Accell siRNA application protocol simplifies targeted gene knockdown

(1) Combine Accell siRNA with Accell delivery media (or other low- or no-serum media). (2) Add Accell delivery mix directly to cells and incubate for 72 hours.


Accell Delivery and Gene Silencing in Cardiomyocytes

Accell Delivery and Gene Silencing in Cardiomyocytes

Accell Delivery and Gene Silencing in Cardiomyocytes

Neonatal rat ventricular myocytes were incubated with 1 μM Accell Green (A; Cat# D-001950-01) or Red (B; Cat# D-001960-01) Non-targeting siRNA for 72 hours in Accell delivery media (Cat# B-005000). Nuclei were stained with DAPI (blue). Labeled control uptake showed diffuse cytoplasmic localization in nearly all cells. The bar graph indicates the level of gene silencing achieved with Accell GAPD Control siRNA (Cat# D-001930-03) and Accell GAPD Control Pool (Cat# D-001930-30) control reagents when used with neonatal rat ventricular myocyte (NRVM) media or Accell delivery media. Myocytes were prepared as described in Maass AH & Buvoli M. Cardiomyocyte preparation, culture, and gene transfer. Methods in Molecular Biology 2007;366: 321-30. mRNA expression was determined by QuantiGene branched DNA assay (Panomics).


References

  1. View the published references citing successful Accell siRNA application.

Citations

  1. Accell siRNA reagents in neuronal cells

    A. Vagnoni et al., Calsyntenin-1 mediates axonal transport of the amyloid precursor protein and regulates Aβ production. Human Molecular Genetics. 13, 2845–2854 (2012). [rat primary cortical neurons (E18)]

    S. Suzuki et al., Differential Roles of Epac in Regulating Cell Death in Neuronal and Myocardial Cells. J. Biol. Chem. 285, 24248-24259 (July 2010). [primary mouse cortical neurons (E15-17)]

    A. M. Dolga et al., TNF-alpha-mediates neuroprotection against glutamate-induced excitotoxicity via NF-kappaB-dependent up-regulation of K2.2 channels. J. Neurochem. 107, 1158-1167 (2008). [mouse primary cortical neurons]

    U. Dreses-Werringloer et al., A Polymorphism in CALHM1 Influences Ca2+ Homeostasis, Ab Levels, and Alzheimer’s Disease Risk. Cell. 133, 1149–1161 (27 June 2008). [SHSY-5Y; human neuroblastoma]

    J. Sebo et al., Requirement for Protein Synthesis at Developing Synapses. J. Neurosci. 29, 9778-9793 (2009). [rat primary hippocampal neurons]

    P. Mergenthaler et al., Mitochondrial hexokinase II (HKII) and phosphoprotein enriched in astrocytes (PEA15) form a molecular switch governing cellular fate depending on the metabolic state. PNAS. 109(5), 1518-1523 (31 January 2012). [extended duration silencing in rat primary cortical neurons]

  2. Accell siRNA reagents in immunological cells

    S. Winning et al., Acute Hypoxia Induces HIF-Independent Monocyte Adhesion to Endothelial Cells through Increased Intercellular Adhesion Molecule-1 Expression: The Role of Hypoxic Inhibition of Prolyl Hydroxylase Activity for the Induction of NF-B J. Immunology. 185, 1786 -1793 (July 2010). [THP-1 monocytes]

    V. Saini et al., CXC Chemokine Receptor 4 Is a Cell Surface Receptor for Extracellular Ubiquitin. J. Biol. Chem. 285, 15566 – 15576 (May 2010). [THP-1 monocytes]

    J. W. Perry et al., Endocytosis of Murine Norovirus 1 into Murine Macrophages Is Dependent on Dynamin II and Cholesterol. J. Virol. 84, 6163-6176 (2010). [murine macrophages]

    M. Steenport et al., Matrix Metalloproteinase (MMP)-1 and MMP-3 Induce Macrophage MMP-9: Evidence for the Role of TNF-a and Coclooxygenase-2. J. Immunology. 183(12), 8119-8127 (15 December 2009). [RAW264.7 macrophages, doi:10.4049/jimmunol.0901925]

    C. B. Lai et al., Creation of the two isoforms of rodent NKG2D was driven by a B1 retrotransposon insertion. Nucleic Acids Res. 37(9), 3032-3043 (May 2009). [mouse NK cell line, gkp174]

    N. Mookherjee et al., Intracellular Receptor for Human Host Sefense Peptide LL-37 in Monocytes. J. Immunol. 183, 2688-2696 (2009). [THP-1; human monocytes]

    D. A. Smirnov et al., Genetic Analysis of Radiation-induced Changes in Human Gene Expression. Nature. 459(7246), 587-91.(28 May 2009).  [immortalized B cells, doi:10.1038/nature07940]

    A. M. McElligott et al., The Novel Tubulin-Targeting Agent Pyrrolo-1,5-Benzoxazepine-15 Induces Apoptosis in Poor Prognostic Subgroups of Chronic Lymphocytic Leukemia. Cancer Research. 69(21), 8366-8375. (13 October 2009). [PGA-1; EBV-transformed chronic lymphocyctic leukemia (CLL) B cell line, 10.1158/0008-5472.CAN-09-0131 ]

  3. Accell siRNA reagents in vivo

    H. Nakajima et al., A rapid, targeted, neuron-selective, in vivo knockdown following a single intracerebroventricular injection of a novel chemically modified siRNA in the adult rat brain.  J. Biotechnology. 157(2), 326-333 (January 2012). [brain injection]

    E. Gonzalez-Gonzalez et al., Silencing of Reporter Gene Expression in Skin Using siRNAs and Expression of Plasmid DNA Delivered by a Soluble Protrusion Array Device (PAD). Molecular Therapy. 18(9), 1667-1674 (22 June 2010). [mouse intradermal injection, doi:10.1038/mt.2010.126]

    P. Bonifazi et al., Intranasally Delivered siRNA Targeting PI3K /Akt /mTOR Inflammatory Pathways Protects from Aspergillosis. Mucosal Immuno. 3(2), 193-205 (18 November 2009). [in vivo intranasal delivery, doi:10.1038/mi.2009.130]

    A. DiFeo et al., KLF6-SV1 Is a Novel Antiapoptotic Protein That Targets the BH3-Only Protein NOXA for Degradation and Whose Inhibition Extends Survival in an Ovarian Cancer Model. Cancer Res. 69, 4733–4741 (2009). [in vivo mouse model]

    Q. Li et al., Silencing MAP Kinase-activated Protein Kinase -2 Arrests Inflammatory Bone Loss. J. Pharmacol. Exp. Ther. 336(3), 633-642 (March 2011). [direct injection to rat gingival tissue; in vivo model of periodontal bone loss]

  4. Accell siRNA reagents in primary, stem, tumor and other cell types

    M. Liao et al., Inhibition of Hepatic Organic Anion-transporting Polypeptide by RNA Interference in Sandwich-cultured Human Hepatocytes: An in vitro Model to Assess Transporter-mediated Drug-drug Interactions. Drug Metabolism and Deposition. 38(9), 1612-1622 (August 2010). [freshly isolated human hepatocytes]

    S. Byas et al., Human Embryonic Stem Cells Maintain Pluripotency after E-Cadherin Expression Knockdown. FASEB J. 24, lb172 (April 2010). [H9 stem cell lines]

    S. Desai et al., PRDM1 Is Required for Mantle Cell Lymphoma Response to Bortezomib Mol. Cancer Res. 8, 907-918 (June 2010).

    I. Barbieri et al., Constitutively Active Stat3 Enhances Neu-Mediated Migration and Metastasis in Mammary Tumors via Upregulation of Cten. Cancer Res. 70, 2558-2567 (March 2010).

    C. Bartholomeusz et al., PEA-15 Induces Autophagy in Human Ovarian Cancer Cells and is Associated with Prolonged Overall Survival.  Cancer Res. 68, 9302-9310 (2008). [OVCA 420; ovarian carcinoma]

    B. Tunquist et al., Mcl-1 Stability Determines Mitotic Cell Fate of Human Multiple Myeloma Tumor Cells Treated with the Kinesin Spindle Protein Inhibitor ARRY-520. Mol. Cancer Ther. 9, 2046–2056 (July 2010). [multiple myeloma cell lines]

    M. Chetane et al., Interleukin-7 mediates glucose utilization in lymphocytes through transcriptional regulation of the hexokinase II gene.Am. J. Physiol. Cell. Physiol. 298(6), C1560-C1571 (Jun 2010). [lymphocytes]

    G. A. Peters et al., The double-stranded RNA-binding protein, PACT, is required for postnatal anterior pituitary proliferation. PNAS. 106(26), 10696-10701 (30 June 2009). [GH3; rat somatolactotrophs (pituitary cell line) and LßT2 gonadotrophs; basophilic cell of the anterior pituitary]