siRNA

esiRNA/esiFLEX/Non-coding

MISSION esiRNA

Eupheria Biotech

 



On this page:
MISSION esiRNA
esiRNA Targeting Long Non-coding RNA
esiRNA Screening Video Tutorial
esiRNA FAQ (Frequently Asked Questions)
Benefits
MISSION esiRNA Offering
Ordering Information
esiRNA Controls
References

MISSION® esiRNA

MISSION esiRNA provides RNAi researchers with a proven, cost-effective, and simple way to perform RNAi screens. MISSION esiRNAs are endoribonuclease-prepared siRNA pools comprised of a heterogeneous mixture of siRNAs that all target the same mRNA sequence. These multiple silencing triggers lead to highly specific and effective gene silencing with lower off-target effects than single or pooled siRNAs.

Product Offerings

Individual esiRNA — esiRNAs provided in tubes at either 20 ug or 50 ug quantities targeting human or mouse genes

esiFLEX esiRNA — esiRNAs provided in 96-well customized plates at either 1 ug, 2.5 µg or 5.0 ug targeting human or mouse genes

Plated esiRNA — esiRNAs provided in 96-well customized plates at either 20 ug or 50 ug quantities targeting human or mouse genes

Genome Scale esiRNA Libraries — Two Genome-scale collections of individual esiRNAs including 16,744 human or 14,068 mouse esiRNAs at either 1 ug, 2.5 ug or 5.0 µg quantities in 384-well plates

Long Non-Coding RNA esiRNA Libraries — Two esiRNA libraries targeting non-coding transcripts including 643 esiRNAs from the mouse genome and 1,245 esiRNAs from the human genome..

esiOPEN esiRNA — Custom esiRNAs targeting the species specific 500 base pair gene sequence provided, supplied in tubes at either 20 μg or 50 μg quantities

esiSEC esiRNA — Secondary independent esiRNA for any human and mouse primary esiRNA, supplied in tubes at either 20 μg or 50 μg quantities

How are MISSION esiRNAs prepared?

esiRNA or Endoribonuclease-prepared siRNAs are pools of siRNAs resulting from cleavage of long double-stranded RNA (dsRNA) with Escherichia coli RNase III. Please see Figure 1.

Figure 1: Overview of MISSION esiRNA Production.
Note: This is for the production of an individual esiRNA
(click image to enlarge).

Generation of PCR products

The template for in vitro transcription is generated by PCR amplification of the cDNA from the clone using primers specific for the vector backbone or target-specific primers appended with RNA-Polymerase promoter sequences. The PCR product is sequence verified. The amplified region is selected based on the highest possible number of highly effective siRNA based on the DEQOR siRNA design program. MISSION esiRNA are designed to cover all known transcript variants of the target gene.

In vitro transcription and the production of long dsRNA

The PCR product generated from the cDNA is used for in vitro transcription reactions. The dsRNA is produced utilizing RNA Polymerases and the two single RNA strands are subsequently annealed before digestion.

Enzymatic digestion

The long dsRNA is enzymatically digested to short dsRNAs. This digestion produces complex pools of siRNA-like molecules.

Purification of digested dsRNA

The digestion is then purified to remove any remaining DNA template, unincorporated nucleotides, and dsRNAs longer than approximately 40 bp. This purification step results in MISSION esiRNAs with an average length of 21 bp.

MISSION esiRNAs

Since the starting material is cDNA, each MISSION esiRNA is guaranteed to target a real gene, no more wasted time on assays for "predicted" genes that do not actually exist.

MISSION esiRNA are a heterogeneous mixture of siRNAs that all target the same mRNA sequence. These multiple silencing triggers lead to highly specific and effective gene silencing.

esiRNA video tutorial for RNAi Screening in Mammalian Cells
esiRNA Video

esiRNA FAQ (Frequently Asked Questions)




Spend your time on discovery!

Benefits

esiFlex™ Woman

  • Lower off target effects than single or pooled siRNAs
  • Guaranteed ≥70% knockdown at the mRNA level
  • Lowest cost genome scale RNAi screening tool
  • >16,000 human and >14,000 mouse targets available
  • Provides quick primary screens




MISSION esiRNA Offering

MISSION esiRNA and MISSION esiFLEX are available individually, as custom arrayed panels, or genome scale libraries that target both human and mouse genes.

 

  Individual esiRNAs Plated MISSION esiRNAs esiRNA Genome Scale Libraries esiFLEX esiRNAs
Quantities available 20 ug
50 ug
20 ug
50 ug
1 ug
2.5 ug
5.0 ug
1 ug
2.5 ug
5.0 ug
Gene target sources
available
Human
Mouse
Human
Mouse
Human
Mouse
Human
Mouse
Packaging 2 ml tubes 96-well plate
(customizable)
384-well plate 96-well plate
(customizable)

 

  esiOPEN esiSEC Non-Coding esiRNA Libraries
Quantities available 20 ug
50 ug
20 ug
50 ug
1 ug
2.5 ug
5.0 ug
Gene target sources available All species Human
Mouse
Human
Mouse
Packaging 2 ml tubes 2 ml tubes 96-well plate

Click here to view additional esiRNA product information.
Browse a complete list of esiRNA.

Individual esiRNA

Our individual esiRNAs are ideal for the biological characterization of single genes. Currently, we offer esiRNAs targeting 16.744 human and 14.068 mouse transcripts at quantities of either 20 µg or 50 µg normalized to 200 ng/µl; larger scales are available upon inquiry. All esiRNAs are designed based on sequence annotations from ENSEMBL database and packaged in single tubes.

Plated esiRNA

esiRNAs provided in 96-well customized plates at quantities of either 20 ug or 50 ug normalized to 200 ng/µl. Currently, we offer esiRNAs targeting 16.744 human and 14,068 mouse transcripts. Plate orders must have ≥10 targets.

Genome Scale esiRNA Libraries

Our genome scale esiRNA libraries are state-of-the-art RNAi reagents especially suitable for large-scale loss-of-function screening. Genome-scale collections of individual esiRNAs for human or mouse species are available. Currently, we offer a human library composed of 16.744 esiRNA and a mouse library with 14.068 esiRNAs. All esiRNAs are designed based on sequence annotations from the ENSEMBL database. The libraries are delivered in 384-well plates with all positions occupied at either 1 µg, 2.5 µg or 5.0 µg quantities normalized to 50 ng/µl. Other quantities, custom arrays or other plate formats are available upon inquiry.

esiFLEX esiRNA

Our esiFLEX sub-genomic collections of esiRNAs provide a convenient small scale option with plate position customization. We provide esiFLEX for human or mouse species and offer 16,744 human and 14,068 mouse esiRNAs in our portfolio. The esiFLEX libraries are provided in 96-well plates at either 1 µg, 2.5 µg or 2.5 µg quantities normalized to 50 ng/µl. The position of each esiRNA on the plates can freely be chosen. Other quantities, concentrations or plate formats are available upon inquiry. All esiRNAs are designed based on sequence annotations from the ENSEMBL database. Minimum order of 24 targets required.

esiOPEN esiRNA

esiOPEN provides access to fully flexible custom-targeting esiRNAs. Provide any transcript-sequence (minimum 500 bp) regardless of the species and an esiRNA will be synthesized against the best target-region (determined by DEQOR) that lies within the provided sequence to ensure maximum efficiency and specificity. esiOPEN esiRNAs are offered at either 20 µg or 50 µg scales normalized to 200 ng/µl and packaged in single tubes. Larger scales are available upon inquiry.

esiSEC esiRNA

esiSEC provides secondary independent silencing triggers for any primary esiRNA in our portfolio. esiSEC are used for the validation of RNAi-phenotypes induced by our primary esiRNAs. esiSEC esiRNAs are offered at either 20 µg or 50 µg scales normalized to 200 ng/µl and packaged in single tubes. Larger scales are available upon inquiry. All esiRNAs are designed based on sequence annotations from the ENSEMBL database.


esiRNA Ordering Information

Individual esiRNAs
Easily find your MISSION esiRNA target by inserting your gene of interest into the search box above and add selected esiRNA to your cart.

For all other esiRNA product offerings:

Controls

  • Negative controls: RLUC, FLUC, and eGFP MISSION esiRNAs
  • Positive control: eg5 (KIF11) MISSION esiRNA (strong mitotic arrest/viability phenotype)
  • MISSION esiRNA targeting eGFP can also be used as a positive control for knockdown in cells expressing eGFP
  • Validated human MISSION esiRNA—Many common gene targets, including LAMIN A, PLK4, and AURKB, have been validated for >70% mRNA knockdown. Validated MISSION esiRNA are suitable for transfection optimization and as positive controls
  • qPCR validation data
  • Western Blot validation data


Validated Human MISSION esiRNA
Gene Name esiRNA Product No. Gene Name esiRNA Product No. Gene Name esiRNA Product No.
ANAPC1 EHU029991 DCTN2 EHU086581 PLK4 EHU001691
ANAPC10 EHU108801 ETN3 EHU040581 RAD21 EHU108911
ANAPC5 EHU069691 H3F3A EHU127091 RCD-8 EHU094431
AURKB EHU001471 INCENP EHU001891 SEC13L1 EHU152131
c14orf10 EHU143981 KIF11 EHU019931 SEH1L EHU049161
CENPA EHU098081 KIF23 EHU066121 SUV39H2 EHU093651
CEP192 EHU026711 LMNA EHU063791 TACC3 EHU063201
CETN2 EHU137031 MAD2L1 EHU074611 TUBG1 EHU056411
ch-TOG EHU078221 NEK2 EHU109951 TUBGCP3 EHU073291
CLASP1 EHU028941 NUMA EHU059141 TPX2 EHU093011
CP110 EHU007261 NUP37 EHU048011    



References

MISSION esiRNA — Proven RNAi Screening Tool

Roguev, A. et al. Quantitative genetic-interaction mapping in mammalian cells. Nature Methods 10, 432–437 (2013).

Abbasi, M., Lavasanifar, A. & Uludag, H. Recent attempts at RNAi-mediated P-glycoprotein downregulation for reversal of multidrug resistance in cancer. Medicinal research reviews 33, 33-53 (2013).

Alvarez-Calderon, F., Gregory, MA. & DeGregori, J. Using functional genomics to overcome therapeutic resistance in hematological malignancies. Immunologic research 55, 100-15 (2013).

Kim, N. & Song, K. KIFC1 is essential for bipolar spindle formation and genomic stability in the primary human fibroblast IMR-90 cell. Cell structure and function (2013).

Ngondo-Mbongo, RP., Myslinski, E., Aster, JC. & Carbon, P. Modulation of gene expression via overlapping binding sites exerted by ZNF143, Notch1 and THAP11. Nucleic acids research (2013).

Vella, P. et al. Tet Proteins Connect the O-Linked N-acetylglucosamine Transferase Ogt to Chromatin in Embryonic Stem Cells. Molecular cell 49, 645-56 (2013).

Calegari, F. et al. Tissue-specific RNA interference in postimplantation mouse embryos with endoribonuclease-prepared short interfering RNA. PNAS, 99, 14236-14240 (2002).

Lin, Z. et al. (2011) Reduced Level of Ribonucleotide Reductase R2 Subunits Increases Dependence on Homologous Recombination Repair of Cisplatin-Induced DNA Damage. Mol Pharmacol doi:10.1124/mol.111.074708. Abstract

Krastev, D.B. et al. A systematic RNAi synthetic interaction screen reveals a link between p53 and snoRNP assembly. Nat Cell Biol 13, 809-18 (2011). Abstract

Nitzsche, A. et al. RAD21 cooperates with pluripotency transcription factors in the maintenance of embryonic stem cell identity. PLoS One 6, e19470 (2011). Abstract

Leushacke, M. et al. An RNA interference phenotypic screen identifies a role for FGF signals in colon cancer progression. PLoS One (2011), accepted for publication.

Slabicki M., et al. (2010) A Genome-Scale DNA Repair RNAi Screen Identifies SPG48 as a Novel Gene Associated with Hereditary Spastic Paraplegia. PLoS Biol 8(6): e1000408. doi:10.1371/journal.pbio.1000408. Paper (2.22 Mb PDF)

Collinet, et al. Systems survey of endocytosis by multiparametric image analysis. Nature 464, 243-249 (2010). Abstract

Theis, M. et al. Comparative profiling identifies C13orf3 as a component of the Ska complex required for mammalian cell division. EMBO J. 28, 1453-65 (2009). Abstract

Ding, L. et al. A Genome-Scale RNAi Screen for Oct4 Modulators Defines a Role of the Paf1 Complex for Embryonic Stem Cell Identity. Cell Stem Cell. 9, 403-15 (2009). Abstract

Kittler, R. et al. Genome-scale RNAi profiling of cell division in human tissue culture cells. Nat. Cell Biol. 9, 1401-12 (2007). Abstract

Kittler, R. et al. An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division. Nature 432, 1036-40 (2004). Abstract

Additional esiRNA Literature References

Stewart, G. S. et al. The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. Cell 136, 420-34 (2009). Abstract

Lawo, S. et al. HAUS, the 8-Subunit Human Augmin Complex, Regulates Centrosome and Spindle Integrity. Curr Biol., 19, 816-26 (2009). Abstract

Fazzio, T. G. et al. An RNAi screen of chromatin proteins identifies Tip60-p400 as a regulator of embryonic stem cell identity. Cell 2008 134, 162-74 (2008). Abstract

Konstantinova, I. et al. EphA-Ephrin-A-mediated beta cell communication regulates insulin secretion from pancreatic islets. Cell 129, 359-70 (2007). Abstract

Nikolova, G. et al. The vascular basement membrane: a niche for insulin gene expression and Beta cell proliferation. Dev Cell. 10, 397-405 (2006). Abstract

Kittler, R. et. al. RNA interference rescue by bacterial artificial chromosome transgenesis in mammalian tissue culture cells. Proc. Natl. Acad. Sci. U.S.A. 102, 2396-401 (2005). Abstract

Liu, W. Y. et al. Efficient RNA interference in zebrafish embryos using synthesized with SP6 RNA polymerase. Dev. Growth Differ. 47(5), 323-31 (2005). Abstract

Yang, D. et al. Short RNA duplexes produced by hydrolysis with Escherichia coli RNase III mediate effective RNA interference in mammalian cells. Proc. Natl. Acad. Sci. U.S.A. 99, 9942-7 (2002). Abstract

 


Contact Us

For technical questions concerning esiRNAs, please email our technical Service department at techserv@sial.com

MISSION is a registered trademark of Sigma-Aldrich Co. LLC Label License.

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