- Synthetic miRNA mimics for miRNA functional analyses and miRNA target site validation
- Optimized design results in correct strand utilization by RISC
- Pre-designed molecules for miRNAs listed in miRBase—custom design available
- Available individually or in sets for miRNA screening
- Purified and ready to transfect
Pre-miR™ miRNA Precursor Molecules are small, chemically modified double-stranded RNA molecules designed to mimic endogenous mature miRNAs. These ready-to-use miRNA mimics, which are similar to, but not identical to siRNAs, can be introduced into cells using transfection or electroporation parameters similar to those used for siRNAs. They enable detailed study of miRNA biological effects via gain-of-function experiments (Figure 1). Positive and negative control Pre-miR miRNA Precursors (listed above) or the Pre-miR™ Precursor Starter Kit can be used to optimize transfection of Pre-miR miRNA Precursors.
Pre-miR miRNA Precursor Molecules have been carefully designed and modified to ensure that the correct strand, representing the desired mature miRNA, is taken up into the RNA-induced silencing-like complex responsible for miRNA activity (Figure 2). In contrast to miRNA expression vectors, these synthetic molecules can be used in dose response studies due to their introduction directly into the cell by transfection or electroporation. Thanks to their small size, they are easier to transfect than vectors, and can be delivered using conditions identical to those used for siRNAs.
Use of the Pre-miR miRNA Precursors enable miRNA functional analysis by up-regulation of miRNA activity. Specific experimental designs include:
- miRNA target site identification and validation
- Screening for miRNAs that regulate the expression of a gene
- Screening for miRNAs that affect a cellular process
miRNA-related cellular functions identified through use of Pre-miR miRNA Precursors can be further investigated using Ambion's Anti-miR™ miRNA Inhibitors, which enable loss-of-function experiments through reduction of miRNA activity.
>> Find & Order Pre-miR miRNA Precursors for your miRNAs of interest through our GeneAssist™ miRNA Workflow Builder.
NOTE: Pre-miR miRNA Precursors are not hairpin constructs and should not be confused with pre-miRNAs.
Pre-miR™ hsa-miR-1 miRNA Precursor Positive Control
Ambion offers the Pre-miR™ hsa-miR-1 miRNA Precursor for use as a positive control in experiments utilizing Pre-miR miRNA Precursors. This precursor was chosen as a positive control because, upon delivery into cells, it effectively down regulates the expression of twinfilin-1, also known as PTK9, at the mRNA level (60-95% reduction; see Figures 3 & 4 at right). Effective delivery and activity of the Pre-miR hsa-miR-1 miRNA Precursor can be detected by real-time PCR using a TaqMan® Gene Expression Assay to PTK9 (Assay IDs: Hs00702289_s1 [human], Mm01598980_g1 [mouse]). Find assay on Applied Biosystems website.
Pre-miR™ miRNA Precursor Negative Controls
Pre-miR™ Negative Controls are random sequence Pre-miR molecules that have been extensively tested in human cell lines and tissues and validated to not produce identifiable effects on known miRNA function. Cy™3 and FAM™ dye-labeled Pre-miR Negative Controls are also available for monitoring transfection efficiency in transfection experiments using Pre-miR miRNA Precursors. The fluorescent label enables direct observation of the cellular uptake, distribution, and localization of the control. These dye-labeled controls are labeled at their 5' end and have the same oligonucleotide sequence as unlabeled Negative Control #1.
Pre-miR™ miRNA Precursor Libraries
The Pre-miR™ miRNA Precursor Library-Human V3 consists of 470 miRNA mimics corresponding to 470 human mature miRNAs cataloged in version 9.2 of the miRBase Sequence Database. This collection of miRNA mimics enables rapid study of miRNA function (Figure 3) and can be used to simplify identification of miRNAs that interact with a particular target transcript. Each Pre-miR miRNA Precursor is in a quantity of 0.25 nmol, dried in individual wells of a 96 well plate. This amount is sufficient for 250 transfections when used at 10 nM in 96 well plates.
[View Pre-miR Library-Human V3 Annotation File]
The Pre-miR™ miRNA Precursor Library–Mouse V3 consists of 379 miRNA mimics corresponding to 379 mouse mature miRNAs cataloged in version 9.2 of the miRBase Sequence Database. This collection of miRNA mimics enables rapid study of miRNA function and can be used to simplify identification of miRNAs that interact with a particular target transcript. Each Pre-miR miRNA Precursor is supplied dried at 0.25 nmol in individual wells of a 96 well plate. This amount is sufficient for 250 transfections when used at 10 nM in 96 well plates.
[View Pre-miR Library-Mouse V3 Annotation File]
Custom Pre-miR miRNA Precursor Libraries, which include 50 or more Pre-miR miRNA Precursors of your choice in the amount you specify, are also available. Contact us at libraries@ambion.com for additional details.
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