Accell siRNA Reagents - Mouse

Targeted gene silencing in difficult-to-transfect cells


A novel siRNA for difficult-to-transfect cells, Accell siRNA is modified to require no transfection reagent or viral vector for delivery. Available as genome-wide, predesigned SMARTpool and single siRNA reagents for human, mouse, and rat.

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The purchase of a Set of 4 Upgrade requires a prior or concurrent purchase of the SMARTpool reagent corresponding to the same gene. Order history will be reviewed for compliance. Questions? Please contact Customer Service.

Enter a common gene identifier and click SEARCH to find products for your gene.

 
Accell SMARTpool - Mouse
E-MOUSE-XX-0005 5 nmol - $635.00select
Accell Set of 4 - Mouse
EQ-MOUSE-XX-0002 2 nmol - $995.00select
Accell individual siRNA - Mouse
A-MOUSE-XX-0005 5 nmol - $320.00select
Accell Set of 4 Upgrade - Mouse
EU-MOUSE-XX-0002 2 nmol - $584.00select

Accell siRNA achieves what no other RNAi product can claim: delivery into difficult-to-transfect cells without transfection reagents, virus, or instrumentation. 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.

Accell siRNA is available as predesigned, genome-wide SMARTpool and single siRNA reagents. Simply search for your gene of interest and choose from the available product formats and quantities.

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
  • Proven performance in neuronal, immunological, primary, and other difficult-to-transfect cell types
  • Appropriate for use in vivo, due to stabilizing modifications
  • Extended-duration knockdown with optimized continuous application

Experimental considerations

This breakthrough in siRNA delivery requires no transfection reagent, but has some unique application requirements.

  • 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.

Product Formats

  • SMARTpool: A mixture of 4 siRNA provided as a single reagent; providing advantages in both potency and specificity
  • Set of 4: A convenient option for purchasing aliquots of all 4 individual siRNAs targeting a single gene
  • Set of 4 Upgrade: Discounted price on the Set of 4 based on a concurrent or prior purchase of the SMARTpool reagent for the same gene
  • Individual siRNAs: Select 1, 2, 3, or 4 individual siRNAs per gene

Sequence information is provided with purchase of pre-designed siRNA products.

  
HazardousNo
Shipping ConditionAmbient
Stability at Recommended Storage ConditionsAt least 12 months
Storage Condition-20 C

Ordering Guidelines

  Approximate # reactions (wells) at 1 µM siRNA concentration (assuming no loss from pipetting)
nmol 96-well plate
(100 µL total reaction volume)
24-well plate
(500 µL total reaction volume)
12-well plate
(1000 µL total reaction volume)
1 10 2 1
2 20 4 2
5 50 10 5
10 100 20 10

Due to the unique nature of Accell siRNA delivery, it requires a higher working concentration than conventional siRNAs. The table provides the approximate number of reactions (wells) at recommended 1 μM Accell siRNA working concentration in different plate formats.

Accell siRNA demonstrates effective uptake and silencing in organotypic brain slices

Accell siRNA demonstrates effective uptake and silencing in organotypic brain slices

Accell siRNA demonstrates effective uptake and silencing in organotypic brain slices

Accell siRNA shows increased uptake with extended incubation. 250 μm cerebellar sections were prepared, cultured, and incubated for 3 hours (A) and 72 hours (B) with Accell Red Non-targeting control (NTC) siRNA before inspection by microscopy.


Accell siRNA effectively silences target genes in cultured P8 mouse brain slices

Accell siRNA effectively silences target genes in cultured P8 mouse brain slices

Accell siRNA effectively silences target genes in cultured P8 mouse brain slices

Cultured brain slices were assessed following incubation with Accell siRNA targeting PPIB and GAPDH or Non-targeting controls (NTC) at three time points (48, 72, 96 hours) and the level of remaining protein was evaluated using Western blot analysis (Actin as loading control).


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.


Accell siRNA delivery and gene silencing in cardiomyocytes

Accell siRNA delivery and gene silencing in cardiomyocytes

Accell siRNA 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 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).


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.


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. 21, 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 (Jul 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(4), 1158-1167 (November 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(7), 1149-1161 (27 June 2008). [SHSY-5Y; human neuroblastoma]

    J. Sebo et al., Requirement for Protein Synthesis at Developing Synapses. J. Neurosci. 29(31), 9778-9793 (5 August 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 USA. 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(20), 15566-15576 (14 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(12), 6163-6176 (June 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. (2009). [mouse NK cell line, gkp174]

    N. Mookherjee et al., Intracellular Receptor for Human Host Sefense Peptide LL-37 in Monocytes. J. Immunol. 183(4), 2688-2696 (15 August 2009). [THP-1; human monocytes]

    D. Smirnov et al., Genetic Analysis of Radiation-induced Changes in Human Gene Expression. Nature. 459(7246), 587-591 (28 May 2009). [immortalized B cells]

    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 (20 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 (September 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) [doi:10.1038/mi.2009.130, in vivo intranasal delivery]

    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(11), 4733–4741 (12 May 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]