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Custom miRNA Inhibitor Synthesis


MicroRNA (miRNA) are small, non-coding RNAs that mediate post-transcriptional gene regulation through binding to complementary sequences of target mRNAs. The miRNA inhibitors, also called antagomirs or antimiRs, are a type of molecules that have the ability to block miRNA regulation of target gene expression. They are designed and optimized for miRNA loss of function research.

With advanced technology and specialized infrastructure, Creative Biolabs offers a full range of custom miRNA inhibitor services as synthetic oligonucleotides and vector-based expression clones for biotechnology, pharmaceutical, and related industries. Meanwhile, we’d like to exactly develop miRNA modulation tools that clients need and design customized inhibitors available for miRNAs not listed in the general database.

Introduction to miRNA Inhibitor

Pathways for mammalian miRNA biogenesis, synthetic RNAi trigger processing and RNAi silencing. Figure 1. Pathways for mammalian miRNA biogenesis, synthetic RNAi trigger processing and RNAi silencing. (Setten, 2019)

miRNAs are 21-25 nucleotide fragments that can negatively regulate gene expression through translational repression or cleavage of a target mRNA. The recognition of natural miRNA targets is primarily determined by perfect complementarity in a seed region (nucleotide positions 2 to 7) with additional interactions contributing in a sequence-specific manner. As for miRNA inhibitors, they are small, synthetically derived molecules, which harness sequence complementary to target mature miRNAs. They can sequester target miRNAs and have already been used to suppress the expression of miRNA targets. Generally, synthetic miRNA inhibitor designs consider the reverse complement of mature miRNAs (target site) and are chemically modified to enhance binding affinity, prevent cleavage of RNA-induced silencing complex (RISC), and give rise to resistance to nucleolytic degradation. When transformed into a cell, binding of endogenous mature miRNAs to these complementary synthetic target sites is thought to be irreversible, therefore, these inhibitors are supposed to seclude endogenous miRNAs, making it unavailable for normal expressions.

Custom miRNA Inhibitor Services at Creative Biolabs

At Creative Biolabs, we provide custom services for miRNA inhibitors using a proprietary algorithm and mature miRNA sequence information. Backed by extensive miRNA expertise and high-quality RNA products, our technical platforms ensure frequent communication, high success rates, and fast timelines during whole projects. Meanwhile, all clients can directly order custom miRNA inhibitor online or submit a detailed request to our technical groups, as well as modify an existing miRNA inhibitor for novel targets and special applications.

Custom miRNA Inhibitor Synthesis

Rational design

We hold a proprietary algorithm and huge mature miRNA sequence data for excellent miRNA inhibitor constructs. The algorithm calculates all possible sequence parameters, selects one predicted to best maintain the synthetic tough decoy (sTuD) structure, offering maximum miRNA recognition and binding. Each sTuD is stabilized by 2’-O-methylated oligos, allowing two miRNA binding sites. As such, the predesigned and designed miRNA inhibitors can regulate gene expression by binding to specific miRNAs and inhibiting their expressions.

Synthesized miRNA inhibitors

Our miRNA inhibitors are chemically synthesized, single-stranded, modified RNAs that can specifically prevent endogenous miRNA activity after transfection into cells. These molecules combine chemical modifications and a totally new secondary structure motif giving improved potency and longevity. We enable clients to order custom-designed miRNA inhibitors not only at cell-culture grade or animal grade, but also available in phosphorothioate modification at different scales (nmol).

Vector-based inhibitors

We have vector-based expression clones both in lentiviral and non-viral vectors for miRNA inhibitors. The inhibitor clones bind to their target miRNAs providing transient and stable suppression of the target genes. The choice of H1 or U6 promoter permits the basic expression of inhibitors in almost all types of mammalian cells. Based on our third-generation lentivector technology, lentivectors express an shRNA that preferentially produces an anti-sense miRNA after processing. The hairpin is rationally designed to be asymmetric, enabling that the sense strand doesn’t have an endogenous miRNA sequence and accomplishing the accumulation of anti-miRNA.

The Items Need to Identify

Target miRNA Information Identify your sequences
Species Identify human, mouse, rat, or other species (inquire)
Modifications Identify the required modifications, such as phosphorylation, 2'-OMe-RNA, etc.
Quantities Identify required quantities, such as 50 nmol, 100 nmol, or 200 nmol.

Highlights

As synthetic miRNA target analogs, miRNA inhibitors are fully complementary, chemically modified oligonucleotides, that have been successfully used to inhibit miRNA function. For all human mature miRNAs and other species miRNAs in the database, Creative Biolabs has reliable design technologies and strategies to produce single-stranded synthetic miRNA inhibitors targeting these miRNAs. Furthermore, we also provide two negative control miRNA to distinguish sequence-specific silencing from non-specific effects. If you don't find what you need on this webpage, please don’t hesitate to contact us for more information.

Reference

  1. Setten, R.L.; et al. (2019). The current state and future directions of RNAi-based therapeutics. Nat Rev Drug Discov. 18(6): 421-446.

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