miRNAs are small non-coding RNA molecules that regulate up to 30% of mammalian gene expression. They serve as post-transcriptional regulators and bind to complementary sequences on target messenger RNA transcripts (mRNAs), resulting in translational repression or target degradation. So far, more than 400 miRNAs have been identified in the human genome and many of them are only different in one or a few nucleotides. Dysregulation of miRNA can be associated with many diseases such as cancer; therefore, the research of miRNAs is very important.

Signosis provides a variety of innovative tools for the research of miRNAs including miRNA arrays for profiling multiple miRNAs simultaneously or monitoring one specific miRNAs.

  

miRNA Real Time PCR

With the miRNA Real-Time PCR kits, miRNA can be analyzed from total RNA or from cell lysates directly without cDNA conversion.

Benefits:

Convenient - All reagents, from cell lysis to PCR, are included in one kit. No RNA preparation - Cell lysates can be used directly in reverse transcription, although total RNA is perfectly fine. Real-time - Analysis can be monitored in real time.

miRNA Northern Blot Assays

The assay kits are non-radioactive with all reagents included, and the probes are biotin pre-labeled. Signosis also provides the Highly Sensitive Northern Blot Assay kits, which are 100 times more sensitive than conventional chemiluminescent detection. Benefits:

No Isotope required - Safe method of detection using chemiluminescence. No probe preparation - Pre-labeled biotin probes included in each kit. All components in one kit- Everything needed to perform the assay is included. Simple procedure - No miRNA isolation required, total RNA can be used directly.

miRNA Luciferase Reporter Vectors

miRNAs have been shown to regulate many developmental and physiological processes, and the deregulation of miRNAs has been linked to a number of disease processes, including cancer. miRNAs play important roles in fine-tuning gene expression regulators referred to as “dimmer switches” because of their ability to repress gene expression without completely silencing it.  The precise monitoring of miRNA expression particularly in cell-based assay, can help to reveal the potential miRNA impacts on cancer etiology, differential stages and therapy development.  Signosis developed the miRNA reporter vectors.  Each reporter vector was constructed by inserting miRNA exactly complementary sequence (~20nt) as corresponding miRNA binding sites on the downstream of luciferase reporter gene.  When miRNA in cells binds to the complementary site at 3'UTR downstream of reporter luciferase gene, it leads to repression of the expression of luciferase and results in reduction of luciferase enzyme activity. The miRNA expression level quantitatively corresponds with the luciferase activities.

 

100+ luciferase reporter vectors are available for monitoring different miRNAs in vivo. Signosis also provides custom services for any unlisted miRNA that you might be interested in.

Benefits:

Functional assay - Monitor miRNA expression in cells. Quantitative analysis - Expression and activation of miRNAs are measured through suppression of firefly luciferase gene expression.

miRNA Luciferase Target Reporter Vectors

 

miRNAs have been shown to regulate many developmental and physiological processes, and the deregulation of miRNAs has been linked to a number of disease processes, including cancer. miRNAs play important roles in fine-tuning gene expression regulators referred to as “dimmer switches” because of their ability to repress gene expression without completely silencing it.  The precise monitoring of miRNA expression particularly in cell-based assay, can help to reveal the potential miRNA impacts on cancer etiology, differential stages and therapy development.  Signosis developed the miRNA target reporter vectors.  Each target reporter vector was constructed by inserting miRNA 3’UTR (~1kb) as corresponding miRNA binding sites on the downstream of luciferase reporter gene.  When miRNA in cells binds to the target site at 3'UTR downstream of reporter luciferase gene, it leads to repression of the expression of luciferase and results in reduction of luciferase enzyme activity. The miRNA expression level quantitatively corresponds with the luciferase activities.

 

 

Benefits:

Functional assay - Monitor the  regulation of target genes by miRNA. Quantitative analysis - Expression and activation of miRNAs are measured through suppression of firefly luciferase gene expression.

miRNA Arrays

The miRNA Arrays are used for simultaneously profiling up to 132 miRNAs.

Benefits:

Highly discriminative power - It is able to differentiate all miRNA isoforms. All let7 isoforms can be clearly distinguished in the assay. Linear amplification - Captured miRNAs are subjected to T7 amplification without introduction of bias in PCR. No pre-isolation of miRNA - Total RNA can be directly used for the assay without pre-isolation of miRNA. Simple procedure - The assay is simple and straightforward.

miRNA Plate Assays

With the Plate Assay kits, the conversion of miRNA into cDNA is not needed. It is 100 times more sensitive than Northern Blot Assay, and the discrimination power of distinguishing the isomers of miRNAs is also higher.

Benefits:

Simple - Do miRNA detection as doing ELISA. It does not need enzymatic conversion from miRNA into cDNA.  The entire assay procedure is “incubation and wash”. Higher sensitivity and better discrimination power - The detection is approximately 100 times more sensitive than miRNA Northern blot.  In addition, it has higher discrimination power than miRNA Northern blot in differentiating miRNA isoforms. Quantitative comparison - The difference of two or more samples can be quantitatively analyzed.

lncRNA Real Time PCR

With the lncRNA Real-Time PCR kits, lncRNA can be analyzed from total RNA .

Benefits:

Convenient - All reagents are included in one kit. Real-time -Quantitative lncRNAs by real-time PCR. Housekeeping gene is provided as positive control.

Direct Hybridization Plate Assays

48 miRNAs can be quantitatively analyzed simultaneously via direct hybridization of RNA sample without any addition steps. The procedure is easy, and the conversion of the miRNA into a cDNA is not needed.

Benefits:

Simple - Do miRNA detection as doing ELISA. It does not need enzymatic conversion from miRNA into cDNA. The entire assay procedure is “incubation and wash”. Higher sensitivity and better discrimination power - The detection is approximately 100 times more sensitive than miRNA Northern blot. In addition, it has higher discrimination power than miRNA Northern blot in differetiating miRNA isoforms. Quantitative comparison - The difference of two or more samples in the expression of a specific miRNA can be quantitatively analyzed.