Mouse Accell 2.0 siRNA Libraries

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Mouse Accell 2.0 siRNA Libraries

Accell 2.0 siRNA libraries are designed to support high-throughput functional gene perturbation in difficult-to-transfect cells.

Using Accell self-delivery chemistry, these libraries enable siRNA screening without transfection reagents, viral vectors, or electroporation, helping streamline workflows in cell types that are often challenging to manipulate.

These siRNA libraries are available in collections spanning the mouse key gene families; and are available in siRNA SMARTpools and set of four individual siRNA formats.

Accell™ 2.0 siRNA Libraries

Accell 2.0 siRNA libraries represent the next generation of self-delivering RNAi reagents, designed for effective gene silencing in difficult-to-transfect cell types without the need for transfection reagents. Sequences are realigned and redesigned according to the latest reference transcriptome, supported by over two decades of siRNA bioinformatic design and RNA chemical synthesis expertise, to support transcriptomic coverage, and target specific gene knockdown across diverse cell types and screening applications.

Collections span the commonly studied gene families, enabling interrogation of hundreds to thousands of genes in both focused and broad discovery screens. Available in 96- or 384-well formats with a choice of siRNA pools or individual unique designs, these libraries support diverse screening workflows and are compatible with single-well analytical techniques including high content imaging, morphological profiling, reporter-based assays, and high-throughput transcriptomics.

Highlights

Next-generation transcriptome alignment and redesign: Accell 2.0 siRNAs are re-annotated and redesigned according to the latest reference transcriptome for improved specificity, with curation to maximize isoform coverage and account for updated genomic coordinates and newly annotated genes.
Recommended for neuronal, suspension, primary, and other difficult-to-transfect cell types.
Proprietary sense strand chemical modification strategy promotes selective antisense/active strand loading into the RISC to reduce off-target effects and facilitates cellular internalization without the need for transfection reagents.
Nuclease resistance modifications protect siRNA integrity in biological environments, supporting sustained gene silencing activity.
Extended-duration knockdown with optimized continuous application.
Available as SMARTpool reagents to minimize seed-mediated off-target signatures while maintaining on-target knockdown.
Available in 96- or 384-well plate formats; Echo® Qualified 384-Well plates available upon request.
Plates include space for control reagent inclusion.
Available as cherry-pick libraries for custom gene selection and hit follow-up.

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

Learn more about Accell 2.0 siRNA design features and benefits on the Accell 2.0 siRNA product page

Available pre-defined collections and approximate gene counts: (table will be filled when we have the gene-count information)

Mouse Accell 2.0 siRNA libraries	Approximate number of genes
GPCRs	470
Ion Channels	420
Phosphatases	310
Proteases	670
Protein Kinases	760
Ubiquitin Enzymes	750
Transcription Factors	1,560
Drug Targets	3,370
Ubiquitin Conjugation Subset 1	90
Ubiquitin Conjugation Subset 2	110
Ubiquitin Conjugation Subset 3	390
Apoptosis	550
Cell Cycle Regulation	170
Cytokine Receptors	140
Deubiquitinating Enzymes	100
DNA Damage Response	240
Epigenetics	810
Membrane Trafficking	140
Nuclear Receptors	50
Tyrosine Kinases	90

Please note, the whole genome library contains the Druggable Genome collection which contains 8 discrete sub-collections (light blue), and all remaining protein coding genes annotated in the relevant reference genome.

Complete your gene silencing experiment by adding these supporting reagents to your order.

Accell 2.0 siRNA Controls

Validated positive and non-targeting negative controls designed for use with next-generation Accell 2.0 siRNAs. Featuring proprietary Accell chemical modifications supporting self-delivery, nuclease resistance, and reduced off-target effects. Available as pooled or individual siRNAs.

Accell 2.0 controls are recommended for any self-delivering siRNA workflow in difficult-to-transfect cell types including neuronal, suspension, and primary cells. Pooled siRNA controls are recommended when additional off-target reduction is desired and are ideal for use alongside Accell 2.0 SMARTpool reagents. Select species-specific positive controls targeting robustly expressed housekeeping genes to confirm reagent function within your experimental system.

Accell 2.0 Control Kits

Accell 2.0 Control Kits provide species-specific fluorescent and unlabeled positive and negative controls to support optimization of siRNA delivery conditions in difficult-to-transfect cells. Designed to assess the performance of next-generation Accell 2.0 chemistry in neuronal, suspension, primary, and other hard-to-transfect cell types, these kits provide a streamlined solution for assay setup and experimental validation.

Select relevant positive controls to efficiently silence a highly expressed housekeeping gene in your experimental system for confidence in your results.

Accell 2.0 Positive Control Reagents	Species	Catalog Number
Accell 2.0 Cyclophilin B Control siRNA	Human, Mouse, Rat	D2-001920-0Y
Accell 2.0 Cyclophilin B Control siRNA Pool	Human, Mouse, Rat	D2-001920-Y0
Accell 2.0 GAPD Control siRNA	Human, Mouse	D2-001930-0Y
Accell 2.0 GAPD Control siRNA Pool	Human, Mouse	D2-001930-Y0
Accell 2.0 Green Cyclophilin B Control siRNA	Human, Mouse, Rat	D2-001970-0Y
Accell 2.0 Red Cyclophilin B Control siRNA	Human, Mouse, Rat	D2-001975-0Y
Accel 2.0 eGFP Control siRNA	N/A	D2-001940-01
Accell 2.0 eGFP Control siRNA Pool	N/A	D2-001940-10
Accell 2.0 Negative Control Reagents	Species	Catalog Number
Accell 2.0 Non-targeting siRNAs	Human, Mouse, Rat	D2-001910-0X
Accell 2.0 Non-targeting siRNA Pool	Human, Mouse, Rat	D2-001910-10
Accell 2.0 Green Non-targeting siRNA	Human, Mouse, Rat	D2-001950-01
Accell 2.0 Red Non-targeting siRNA	Human, Mouse, Rat	D2-001960-01
Accell 2.0 Control siRNA Kits	Species	Catalog Number
Accell 2.0 Control siRNA Kits (Green)	Human, Mouse	K2-005000-G1-0Y
Accell 2.0 Control siRNA Kits (Red)	Human, Mouse	K2-005000-R1-0Y

Peer-reviewed publications demonstrate effective silencing in numerous cell types
Human Suspension Cell Lines	BDCM, IM-9, Jurkat, K662, THP-1, Jeko-1 and Mino MCL
Human Adherent Cell Lines	A-375, ARPE-19, CaCo-2, CG4, DLD-1, DU-145, EC52, GTM-3, MR32, LAN5, LNCaP, MIA PaCa-2, NCI/ADR-RES, OVCAR-3, OVCA-420, PGA1, PNT2, SH-SY5Y, SK-N-BE and SK-N-SH neuroblastoma, SK-BR3, SKOV3, TE15, U251, U87MG, U937
Mouse Adherent Cell Lines	ES-D3, H9c2, LbT2, LNK, mouse oviduct epithelial cells, mProx24, PC-12
Human Differentiated Stem Cells	Adipocytes and osteoblasts from hMSC
Human Primary Cells	HUVEC, HUASMC, hMSC, PBMC, astrocytes, hypothalamic neurons, T cells, NK cells, macrophages, immortalized B cells, esthesioneuroblastoma cells, hepatocytes, bronchial smooth muscle cells
Mouse and Rat Primary Cells	Cortical neurons, hippocampal neurons, striatal neurons, Naïve CD4+ T cells, LN T cells, D1 T cells, RAW264.7 macrophages, hepatocytes, neonatal ventricular cardiomyocytes, fibroblasts derived from genetically-modified mice, bone marrow-derived DC, NOD CD4+ CD25- splenic cells, NOD BMMs, VSMCs, somatolactotroph GH3, McArdle 7777 cells (hepatoma cell line), osteoclast precursors, calvarial (cranial) organ, pituitary cell types derived from primary tumors
In vivo Animal Applications	Please contact Scientific Support for guidance

Application of Accell 2.0 siRNA simplifies targeted gene knockdown

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Combine Accell 2.0 siRNA with Accell siRNA delivery media (or other reduced serum media).
Add Accell 2.0 delivery mix directly to cells and incubate for 72 hours.

Accell 2.0 siRNA maintains performance of gene target knockdown set by Accell siRNA in neurons derived from induced WTC-11 iPSC cells

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Target mRNA knockdown in matrix-free 3D spheroids using Accell 2.0 siRNA

Characterization of target mRNA knockdown by Accell and Accell 2.0 siRNAs in cultured neurons. Data are shown relative to neurons treated with non-targeting control (NTC) siRNA. Experimental schematic (A) and relative gene expression in WTC-11-derived neurons (B). Neurons derived from WTC-11 iPSCs were cultured in differentiation medium for 14 days, with daily medium replenishment. On day 14, 50 µL of medium was removed and replaced with 50 µL fresh medium containing 2 µM Accell or Accell 2.0 pooled siRNA designs (4 siRNA designs at 0.5 µM each) or 2 µM Accell PPIB control (Cat# D-001920-01), resulting in a final siRNA concentration of 1 µM in all conditions. An identical concentration of Accell Non-targeting Control siRNA #1 (D-001910-01) was used for each target NTC condition. siRNA mixtures were replenished by half-medium change on differentiation days 16 and 18, for a total of three siRNA additions. Total RNA was extracted on differentiation day 20 for downstream processing. Figure A is created with BioRender.com.

Performance of target mRNA knockdown in matrix-free 3D spheroids is maintained with Accell 2.0 siRNA

Characterization of Accell and Accell 2.0 siRNA-mediated mRNA knockdown in ECM-free HCT-116 spheroids. Control, PTEN (A), and PPP2R1B (B) mRNA expression were quantified in HCT-116 spheroids after 144 hours of siRNA treatment. (C) Dosing timeline for Accell 2.0 siRNA delivery. Spheroids were generated by seeding 3,000 HCT-116 cells into ECM-free, 96-well ultralow attachment round-bottom plates in HCT-116 medium (McCoy’s 5A medium supplemented with 10% FBS and 2 mM L-glutamine). After two days, 50% of the medium was removed and replaced with 2 µM Accell or Accell 2.0 single or pooled siRNA designs (4 siRNAs at 0.5 µM each) for a final concentration of 1 µM siRNA. Matching concentrations of Accell Non-targeting Control siRNA #1 (D-001910-01) or Accell PPIB control siRNA (D-001920-01) were added on the same schedule. After three rounds of siRNA replenishment, spheroids were lysed, total nucleic acid was collected, and RT-qPCR was used to measure target mRNA abundance. Data are shown relative to target expression in spheroids treated with non-targeting control siRNA. Figure C is created with BioRender.com.

Target protein knockdown in matrix-free 3D spheroids using Accell 2.0 siRNA

Characterization of Accell siRNA-mediated cyclophilin B (PPIB) target protein knockdown in ECM-free HCT-116 spheroids. Data are presented relative to expression in spheroids treated with non-targeting control (NTC) siRNA. (A) Western blot and (B) quantification of Cyclophilin B protein expression in HCT-116 spheroids determined from (A) after 144-h siRNA treatment. Each lane is representative of an independent pool of spheroids (N=3). (C) Immunocytochemistry images of serially sectioned HCT-116 spheroids after 144-h siRNA treatment with an Accell PPIB Control Pool or NTC Pool. Spheroids were then collected, fixed in 2% paraformaldehyde, embedded in optimal cutting (O.C.T.) temperature compound, flash frozen, and serially sectioned. Material was incubated with 1:200 dilution of primary antibody against PPIB (anti-cyclophilin B, Abcam ab16045) and secondary incubation with 1:500 dilution of Alexa 488-conjugated Goat-anti-rabbit antibody (Invitrogen A11034) and 1:1000 dilution of Hoechst 33342 (Molecular Probes H-3570). Scale bar = 200 µM.

Accell control 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 (Cat# D-001960-01-05) before inspection by microscopy.

Robust gene target knockdown in primary lung fibroblasts using Accell 2.0 siRNA

IMR-90 human lung fibroblasts (5,000 cells / well) were plated in 96-well plates in IMR-90 media (EMEM supplemented with 10% FBS, L-glutamine and 0.1 mM non-essential amino acids). After adhesion and outgrowth for 16-18 hours, IMR-90 cells were at 80% confluency and experiments were initiated. After media removal, cells were washed with D-PBS and immersed in 100 mL Accell media with 1 mM Accell 2.0 siRNA. After 5 hours of incubation, 100 mL of IMR-90 media with 4% FBS was added to each well to result in 2% FBS. After culturing for 72 hours, total RNA was extracted and used for downstream RT-qPCR analysis of target mRNA abundance from PTEN, PPP2R1B, AKT1, and AKT2 genes relative to Non-targeting Control sample (NTC).