miRCURY LNA miRNA Family Power Inhibitors

Discover unknown miRNA function

miRNA family members are often co-expressed, either from the same pri-miR transcript or from distinct but co-regulated loci. This suggests that miRNAs within a family may share a common regulatory function. Given their identical seed sequence, miRNA family members will have many mRNA targets in common, and they are therefore likely to be functionally redundant. Therefore, analysis of miRNA family function requires simultaneous inhibition of all or several family members. The miRCURY LNA miRNA Family Power Inhibitors allow you to observe robust phenotypes that would be either absent or weakly presented if using inhibitors of individual miRNA family members (see figure The miR-30 family positively regulates hMADS cell adipocyte differentiation). This enables you to discover new, unknown miRNA functions that you may have deduced through careful analysis of miRNA target predictions, but have not been able to prove experimentally.

Design

miRCURY LNA miRNA Family Power Inhibitors are specifically designed to silence entire families of miRNAs, rather than individual family members. Through careful design, we have developed special family inhibitors that knock down all family members, while retaining specificity for the targeted family (see figure Example of miRNA family inhibitor design). This is achieved through the following design criteria:

• Sequence optimization: identification of a target sequence sufficiently short to be shared by family members and sufficiently long to define the family.
• Mixed-base synthesis: when target sequences differ at a single position, we use a mixture of two bases at the corresponding position during the synthesis of the antisense oligonucleotide. This allows us to generate two oligonucleotides in a single oligonucleotide synthesis reaction.
• Mixture of inhibitors: when it is not possible to target a whole family using a single nucleotide synthesis, we provide a mixture of two or more inhibitors that make up the family inhibitor.
• LNA optimization: family inhibitors are short 10–16mers, but enhanced LNA content ensures that they all have the same target affinity (T_m) as our individual miRCURY LNA miRNA inhibitors.

The miRCURY miRNA Family Power Inhibitors have phosphorothioate backbones for enhanced stability and potency. All 43 Family Power Inhibitors are also available in a library plate format (miRCURY LNA miRNA Inhibitor Libraries).

Coverage

We have developed inhibitors for more than 40 human miRNA families that are conserved in mouse. miRCURY LNA miRNA Family Power Inhibitors have been designed only for miRNA families in which the design criteria described above provide an advantage over simply using a mixture of individual inhibitors of all miRNA family members.We define miRNA families as highly related miRNAs that share an identical seed sequence (nucleotides 2–8). This is particularly important for target recognition, although the miR-200 family is one of the few exceptions to this rule.Some very large, miRBase-defined miRNA families, such as the hsa-miR-506, hsa-miR-515 and hsa-miR-548 families, comprise miRNAs with similar sequences but with different seed sequences. These families are not represented in our miRCURY LNA miRNA Family Power Inhibitors.Power Inhibitors are not recommended for use with cells or cell lines derived from muscle or the central nervous system (CNS). These cell types are known to be particularly sensitive to sequence-dependent toxicity of phosphorothioate-modified oligonucleotides.

miRCURY LNA miRNA Family Power Inhibitors are antisense oligonucleotides with perfect sequence complementarity to their targets. When introduced into cells, they sequester the target miRNA in highly stable heteroduplexes, effectively preventing the miRNA from hybridizing with its normal cellular interaction partners. The sequences of the oligonucleotides and their LNA spiking patterns have been carefully designed to achieve uniform high potency, regardless of the GC content of the target. The T_m is normalized to an optimal temperature, and the level of self-complementarity is kept to a minimum.Following resuspension, miCURY LNA miRNA Family Power Inhibitors are introduced into cells with a transfection reagent or via electroporation. Phenotypic effects are at an appropriate time thereafter.

miRCURY LNA miRNA Inhibitors are primarily used miRNA functional studiesby assessing the biologicalconsequences of inhibiting miRNA activity. These effects can be assessed in a variety of ways, including using cellular assays to monitor cell proliferation, celldifferentiation or apoptosis. The effects on gene expression can also be measured at the mRNAor protein level of putative miRNA targets.