A c t i v a t e R N A i w i t h P r e c i s i o n. Specifically elicit RNAi without dsrna. mrna binding region

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1 SM NEW TECHNOLOGY: Specifically elicit i without ds. attracting loop processing stem m binding region Novel Molecule Gene Knockouts Eliminate Off Target Suppression Use in-situ or in-vivo Avoid Interferon Pathways Avoid Endogenous Gene Regulatory Machinery Available for Transient or Stable i Activat e RN Ai wit h Precision

2 Specifically elicit i by cleavage or blocking of your gene without disadvantages of ds. Precision represents a novel i pre-cursor designed as an more specific means of gene modulation than ds. Precision was developed exclusively by Oligoengine scientists and is based on a novel structure called the sp (self-protecting ). This new molecule can be transfected as or expressed by your preferred i plasmid with specificity over ds. Introduction to the benefits of Precision. Introduction Oligoengine s Precision replaces sh and eschews the idea that i requires double-stranded (ds). In fact, advantages of specificity, stability, and adaptability can be achieved by removing ds altogether. When you consider that all ds products are processed by complexes before functioning, it s easy to imagine a more precise molecule to elicit only the i mechanisms required. Precision is such a molecule. It is processed efficiently and specifically as a pre-cursor of the functional molecule in i: the single strand. The benefit of this new design is that the molecule is more specific to your target gene than typical si. Since Precision is not double-stranded, there is no risk of sense-strand based off-target effects typically associated with si or sh 1. Additionally, Precision can be designed with various attribute to drive processing and aboption into targeted mirnp or RISC complexes 2. This allows Precision to replace m cleaving si or function as a translational silencing molecule. Precision is the latest addition to our Advanced i platform. Oligoengine s Advanced i Platform represents the second generation of i aimed at replacing si and sh in applications of Reverse Genomics, Target Validation, Drug Discovery, and even Therapeutics. We invite you to review this novel platform and examine the advantages to your research. Contents Precision: the molecule...3 Precision: vs. ds specificity...4 Precision: processing theory...5 Precision: silencing in Model Genes EFFICACY: Transient - EFFICACY: Stable i: psuper integration Precision: as a BioPharmaceutical...7 Precision: how to get started...8 Precision: order form...9 Precision: commercial licensing...

3 : the molecule Precision functions similiarily to ds by directing to bind with target m via Watson-crick base pairing to guide a particular mechanism. Precision s inhibitory effects can be manipulated by design. Oligoengine classifies these as attributes which can be added to optimize your results. Design attributes elicit processing pathways and recruiting of complexes to direct function and improve overall efficacy. Effective designs of Precision can easily be produced using your Oligoengine3 (OE3) Software. Basic Form: RISC cleaving guidence processing stem m binding region loop In it s native pre-processed stage, the (small stem-loop, ss, small stem-loop) represents the basic precursor format offered by Oligoengine. Attribute A: translational block or SNPactive nucleotide mismatch to m To disable the cleavage mechanisms, mis-match the central bases to your target m. This alone may not enable effective translational blocking, but can be effective at suppressing SNP containing genes. Attribute B: mirnp complex recruiting Drosha cleave site MIR Mimic Precision Drosha domain Effective translational blocking can be enhanced by recruiting designs. Pathways into mirnp via Drosha can enhance such functionality. Attribute C: cellular delivery ligand site A ligand site b Specific Extracellular delivery via membrane binding domains or carrier molecules represents a future direction for transient forms of Precision. Additionally, the serum life-span has been shown to be -fold stronger than si (see page 7).

4 : specificity vs. ds One major advantage that Precision holds over sh (ds) is that it produces a more specific product. This is achieved by utilizing the unique sp precursor design so that cellular processing of the molecule results in a single functional template. This effective strategy removes sense strand based off-target suppression found in all ds products without sacrificing the ability to elicit i. Single-stranded sp Post-processed Precision NO SENSE STRAND Precise Suppression by Precision m targeting region Target Recognition Primary m Target SINGLE TEMPLATE Off-Target Suppression of si and sh Processing ds i results in sense and antisense guided suppression. ds results in Off-Target suppression of genes complimentary to sense. Overall molecule is double-processed enzymatically resulting in decreased efficiency. Resulting double-stranded results in Off-Target suppression of genes complimentary to sense. Dicer processing of hairpin into si is randomized within stem of hairpin. si -Sense -Antisense sh Short-Hairpin Random Hairpin Cleavage Active RISC cdna Target Template Target Recognition Active RISC Primary m Target Cleaved Primary m Off-Target Recognition Target Recognition Unintended m Target Cleaved Unintended m Primary m Target Cleaved Primary m Cleaved Unintended m

5 : processing theory By design, gene modulation techniques can be directed to guide se cleavage or translational blocking mechanisms. These pathways can be elicited by design attributes of the Precision molecule. While dicer may not be required to pre-process Precision, it s currently believed that dicer s involvement enhances processing rates which lead to increased efficacy of Precision. Processing for Gene Suppression This chart provides an overview of dicer pre-processing for either RISC or mirnp complex processing pathways. Precision Processing by complex Single-stranded sp RISC Single-stranded sp mirnp m targeting region m targeting region DICER DICER RISC mirnp AAAAAA m Cleavage m Translational Arrest

6 : silencing model genes To illustrate the basic silencing efficacy of Precision, Oligoengine has produced assays in models of both transient and stable i against cotransfected Luciferase, and endogenous p53. Examples have been made using public versions of i sequences and then compared to Oligoengine optimized versions in p53. Oligoengine will soon publish results for driving silencing mechanisms, large and sensitive protein-to-protein interactions for specificity, and invivo applications. Stably Expressed Luciferase 2ul TKO, 48 hours, 293 cells, picomoles of ef Precision Precision as Inhibition of Luciferase Expression (Transient ) Control 37 Precision 34 Precision Precision + psuper Inhibition of Luciferase (psuper integration) 24 Control Precision Precision A c ti v a t e R NA i wi t h P re ci s i o n +

7 : silencing model genes Endogenous p53 (Transient ) 2ul TKO, 48 hours, 293 cells, nanomolar of each molecule Inhibition of p53 Expression Published p53 i site Precision targets Engogenous Genes Control si Precision Optimizing Precision Inhibition of p53 Expression Oligoengine Optimized Design 98 Control si Precision 34 Serum Stability Precision si 65 Therapeutics TIME

8 Activat e RN Ai wit h Precision IDENTITY: NAME: fax-back quote form Fax it back, we ll take care of the rest! Fax to PHONE: *Guarantee applies only to orders containing 3 molecules targeting a single gene. One of the three molecules is guaranteed to achieve >% of target gene. Full m sequence must be known and used in the design. 3 Precision (): $999 Precision in form for transient use. Oligoengine s i design is free with this offer. Precision design results are documented and *guaranteed >% effective when designed by Oligoengine. GENE OR SEQUENCE: GENE(S): or N..19 SEQUENCE(S): OPTIONS: Negative Controls: 5 nm Mamm-X Scramble ($69) Base-X Scramble Set ($699) Modifications: FAM ($99) Other Notes: *Guarantee applies only to orders containing 3 molecules targeting a single gene. One of the three molecules is guaranteed to achieve >% of target gene. Full m sequence must be known and used in the design. 3 pairs of Precision (DNA): $299 Precision in DNA form for plasmid ligation. Oligoengine s i design is free with this offer. Design results are documented and guaranteed >% effective. GENE OR SEQUENCE: GENE(S): or N..19 SEQUENCE(S): RESTRICTION ENZYME SITES: BglII / HINDIII (default) BglII / XHOI Other System

9 Licenses available exclusively from Oligoengine. For more information and commercial-use licensing terms: Oligoengine. All Rights Reserved.