Oligoengine's Novel RNAi: Removing off-target suppression Oligoengine RNAi Products

H1 Promoter: A distinct Advantage for shRNA and microRNA

The pSUPER suite of vectors offer the freedom of H1 and the power of Pol III for shRNA expression.

H1 vs. U6

A key component of the pSUPER RNAi System is the H1 RNA polymerase III promoter, which drives the endogenous production of shRNA. While other RNAi vector systems employ U6 promoters, research by the Netherlands Cancer Institute shows H1 to have distinct advantages over other pol III promoters.

Advantages of the Pol III Promoter

Unlike the majority of RNA pol III promoters, type III RNA pol III promoters do not require additional sequence elements and therefore are good candidates for in vivo delivery systems of siRNA. The most abundant cellular RNAs transcribed from a type III RNA pol III promoter are the U6 small nuclear RNA (which play a crucial role in the processing of premature RNA); the 7SK RNA (a negative regulator of RNA polymerase II elongation factor TEFb); and H1 RNA (a component of RNAse P).

The H1 Advantage

The potential negative effects of four different RNA polymerase III promoters on the expression of a heterologous gene driven by a polymerase II promoter were evaluated. The human H1and 7SK promoters displayed no non-specific interferences, whereas the murine and to a lesser extent the human U6 promoters highly interfered with the expression of LGFP. The toxicity of the murine U6 promoter has been previously identified.

Additionally, the H1 promoter proved to offer more flexibility for shRNA target design. In most cases, the +1 nucleotide of the U6-like promoters is always guanosine, whereas the +1 for H1 promoters is adenosine. Interestingly, changing the +1 adenosine to uridine, cytidine, or guanosine within H1-expressed stem-loop sequences does not seen to affect gene silencing, indicating that H1 promoters may be more flexible than U6 promoters in regard to +1 sequence changes1.

Therefore, the human H1 promoter was used in subsequent experiments aimed to evaluate the gene silencing potential of the pSUPER RNAi System, with highly successful results2,3.

Stable Suppression by shRNA or microRNA

The pSUPER RNAi System allows researchers to perform long-term experiments to study the effect of blocking the production of specific proteins or microRNA within cells and whole organisms- leading to new therapeutic weapon against many diseases including HIV, cancer and other genetic disorders.

With silencing effects lasting up to two months or more, this new system also unlocks the power of functional genomics for target validation and drug discovery, two of the key components in the gene-to-drug pathway.

The pSUPER Heritage

The original pSUPER vector was first described in a paper published in the prestigious journal Science in April 2002. Such was its impact that an unprecedented 1,500 requests were received from research scientists asking for access to the vector for their own studies - before the vector was even available for sale.

As a result the pSUPER vector is becoming a standard workhorse in laboratories worldwide for shRNA, microRNA, and X-miR MicroRNA repressors, providing insight into the essential genes involved in a wide range of diseases - from neurodegenerative diseases such as Alzheimer's and Parkinson's, through infectious diseases such as HIV and hepatitis, to cancer and other genetically based diseases.

1. Tuschl, T., Nature Biotechnology 20, 446 (2002)
2. T.R. Brummelkamp, R. Bernards, and R Agami, Science 296, 550 (2002).
3. T.R. Brummelkamp, R. Bernards, and R Agami, Cancer Cell 2:3, 243-247 (2002)

Getting Started with pSUPER or pSUPERIOR

The pSUPER RNAi System generates long-term loss-of-function phenotypes in mammalian cell lines by utilizing a novel vector, along with a pair of oligos unique to the target gene that are ligated into the vector, to direct the persistent synthesis of siRNA intracellularly. Follow these guidelines when designing your oligos:


shRNA Design

A guide to design 60-mer shRNA for pSUPER. shRNA is designed automatically in Oligoengine 2.0.

  • shRNA for pSUPER

Targeting Your Gene with Asymmetric shRNA or X-miR

The pSUPER vector requires the insertion of two annealed long-mer oligos. We have pre-designed thousands of Asymmetric shRNA, microRNA clones, or X-miR MicroRNA Repressors. Please email us for shRNA design specific to your project or check online in our store.

Enzyme Options

shRNA, microRNA repressors, or microRNA for pSUPER contain the ends compatible with digestion using either BglII/HindIII (default) or BglII/XhoI.


miRNA Design

pSUPER clones expressing all known mature microRNA are available in our store

  • miRNA for pSUPER

pSUPER.retro and Packaging Cells

Oligoengine recommends the use of the Phi-NX cell line, a 293T-based line developed by the Nolan Lab at Stanford. The Phi-NX cell line is commercially available at AGCC. Alternatively, other cell lines have proven successful with pSUPER.retro. Phe-NX Cell Line

pSUPERIOR and Inducible RNAi

With the use of a TETR binding protein, pSUPERIOR gives you the option of controlling suppression inducibly with Tetracycline or Doxycycline.. pcDNA6/TR Information

Price List.

All pSUPER and pSUPERIOR vectors are available exclusively in circular form.

pSUPER Prices price
VEC-pBS-0002 pSUPER.basic $299
VEC-pBS-0004 pSUPER.neo $449
VEC-pBS-0006 pSUPER.gfp+neo $499
VEC-pBS-0008 pSUPER.puro $449
VEC-pRT-0002 pSUPER.retro.puro $399
VEC-pRT-0004 pSUPER.retro.neo $449
VEC-pRT-0006 pSUPER.retro.gfp+neo $549
VEC-Ind-0002 pSUPERIOR $599
VEC-Ind-0004 pSUPERIOR.neo $549
VEC-Ind-0006 pSUPERIOR.puro $549
VEC-Ind-0008 pSUPERIOR.gfp+neo $599
VEC-Ind-0010 pSUPERIOR.retro.puro $649
VEC-Ind-0012 pSUPERIOR.retro.neo $649
VEC-Ind-0014 pSUPERIOR.retro.gfp+neo $699
VEC-p53 pSUPER.p53 $149

pSUPER Fax-Back Order Form

All pSUPER products are available within Oligoengine 2.0. Alternatively, use the above fax form to place your pSUPER order.

  • pSUPER Order Form

Any Target + pSUPER or pSUPERIOR + 18 Days.

Pick any pSUPER or pSUPERIOR vector, a target gene, and we will send a sequenced clones for $599 each. When you order three, we will guarantee suppression greater than 70%.

Cloning Service Deliverables:

• Clones can express either Asymmetric shRNA, mature microRNA, or X-miR MicroRNA Repressors.

• All clones are sequence confirmed.

• 18 Business Day turnaround.

• Price includes all materials.

• Delivered as miniprep, ~4 µg.

Cloning Fax Form.

  • pSUPER Order Form

Published Papers Using pSUPER.

We are amassing a list of hundreds of published papers which successfully utilize pSUPER vectors.

Get your paper included.

Email customer service with your reference for using pSUPER and we will be happy to cite your work on the site.

pSUPER RNAi System Protocols

Click to download a PDF of the pSUPER or pSUPERIOR protocol.

pSUPER Protocol:

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pSUPER.retro.neo Protocol:

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pSUPER.retro.puro Protocol:

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pSUPERIOR Protocol:

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pSUPERIOR.retro Protocol:

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pSUPER RNAi System Maps

Click to download a PDF of the pSUPER or pSUPERIOR maps.


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pSUPER.puro Map:

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pSUPER.neo Map:

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pSUPER.neo+gfp Map:

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pSUPER.retro.puro Map:

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pSUPER.retro.neo Map:

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pSUPER.retro.neo+gfp Map:

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pSUPERIOR.puro Map:

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pSUPERIOR.neo Map:

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pSUPERIOR.neo+gfp Map:

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pSUPERIOR.retro.puro Map:

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pSUPERIOR.retro.neo Map:

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pSUPERIOR.retro.neo+gfp Map:

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pSUPER RNAi System Sequences

Click to download a text file of the pSUPER or pSUPERIOR sequences.

pSUPER Sequence:

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pSUPER.puro Sequence:

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pSUPER.neo Sequence:

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pSUPER.neo+gfp Sequence:

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pSUPER.retro.puro Sequence:

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pSUPER.retro.puro (Stuffered Version) Sequence:

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pSUPER.retro.neo Sequence:

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pSUPER.retro.neo+gfp Sequence:

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pSUPERIOR Sequence:

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pSUPERIOR.puro Sequence:

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pSUPERIOR.neo Sequence:

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pSUPERIOR.neo+gfp Sequence:

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pSUPERIOR.retro.puro Sequence:

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pSUPERIOR.retro.neo Sequence:

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pSUPERIOR.retro.neo+gfp Sequence:

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Licensing the pSUPER RNAi System.

Now, to better support the needs of our commercial customers, individual pSUPER vectors are available to non-academic/for profit organizations under more flexible and affordable licensing terms.

Academic and Non-Profit Licensing Terms

Researchers at academic and non-profit institutions are granted a limited end-user license with the purchase of each pSUPER vector.

Commercial and For-Profit Licensing Terms

Previously, use of the pSUPER RNAi System by commercial pharma and biotech companies has been subject to a comprehensive licensing program. Now companies can purchase individual pSUPER vectors under a more streamlined product use agreement. Additionally, any products, including cell lines, transgenic animals and therapeutic compounds developed using the pSUPER RNAi System will be considered as licensed products and subject to a sales royalty by Cancer Research Technologies.

For more information, please contact an OligoEngine Customer Service.


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