RT² miRNA PCR Arrays and individual Assays enable you to obtain high-quality and
genome-wide miRNA expression data by doing nothing more than a simple RT-PCR
protocol. SABiosciences' patent-pending miRNA technology ingeniously integrates a
universal tailing and reverse transcription reaction specific for miRNA with the
accurate expression level measurement of distinct miRNA sequences that may only
differ by a single nucleotide base. With this technology, you can easily get a comprehensive survey of miRNA expression in your cell line, tissue, or paraffin-embedded sample of interest.
Micro RNA (miRNA) are tiny,
typically 22 nucleotides. The
explosive research in this
area has established miRNA as
powerful regulators of gene
expression. Micro RNA has been
implicated in diverse areas
such as the immune response,
neural development, DNA
repair, apoptosis, oxidative
stress response and others. If
you are conducting biological
or biomedical research, you
should examine miRNA
expression profiles just as
you would for other key
players in your pathway. Read
more.
MicroRNAs:
How These Small Fragments of RNA are Having a Large Impact
MicroRNAs (miRNAs) were first identified in the study of C. elegans
development fifteen years ago, but have since been found in nearly every species
of plant and animal. The role in embryogenesis of miRNA was just the tip of the
iceberg, as these powerful regulators of gene expression have been implicated in
such diverse areas as the immune response, neural development, DNA repair,
apoptosis, oxidative stress response and others. It is the goal of this
report to give an overview of how miRNA, a new level of genetic regulation, is
important to both basic as well as applied research in the life sciences.
What is miRNA?
Micro RNA are tiny, typically 22 nucleotides in length, and are produced from
larger precursor RNA transcripts (about 70 nucleotides) by enzymes in the
Argonaute family and the RNase III, Dicer. The miRNA is then incorporated into
the RNA-induced silencing complex (RISC) which suppresses the translation or
enhances the degradation of target messenger RNA (mRNA) molecules, thereby
reducing the synthesis of the encoded protein. The miRNA acts by binding to the
3'-untranslated region of the mRNA, making it possible for a single miRNA to
control multiple genes with the same sequence in this region of the mRNA.
In vertebrates, the RISC complex is guided to its mRNA target by the miRNA
strand, which typically base pairs imperfectly to its target in the 3-prime
untranslated region (3'-UTR), signaling the target for translational repression
through unknown mechanisms. More than 500 miRNA sequences have been identified
in humans (http://microrna.sanger.ac.uk/sequences/index.shtml), and each miRNA
is proposed to have hundreds of mRNA targets due to their imperfect base
pairing. Therefore, the bioinformatic prediction that 30% of human genes are
regulated by miRNA can be seen as a reasonable assumption.
Relevance of miRNA to Human Biology
Before the discovery of miRNA, it had been known that a large part of the
genome is not translated into proteins. This so called "junk" DNA was
thought to be evolution's debris with no function. We now realize that a portion
of this non-coding DNA is highly relevant in the regulation of gene expression.
While only about 500 human miRNA sequenes have been identified so far, genomic
computational analysis indicates that as many as 50,000 miRNA may exist in the
human genome, and each may have multiple targets based on similar sequences in
the 3'-UTR of mRNA.
The importance of the miRNA regulatory pathways is underscored by the
impressive list of diseases which have recently been found to be associated with
abnormal miRNA expression.
Cancer: miRNAs have been found to be down-regulated in a number of
tumors, and in some cases, the reintroduction of these miRNA has been shown to
impair the viability of cancer cells. The value of miRNA profiles in tumor
diagnostics is well established. For instance, strong up and down regulations of
16 miRNA sequences have been shown in primary breast tumors, and these markers
may aid in the development of tests for drug-resistance and for treatment
selection. Underlining the important role miRNA plays in oncology is the
formation of several new companies which seek to expand development of miRNA-based
therapeutics.
Age Related Diseases: Evidence is accumulating that many age-related
diseases are associated with a decreased signaling control that occurs in
mid-life. The miRNA controlling such systems as the cell cycle, DNA repair,
oxidative stress responses, and apoptosis have been shown to become abnormally
expressed in midlife. It is highly likely that continued research will reveal
important associations with the aging process, and may lead to therapeutics that
can improve the quality of life.
Heart Disease: Two heart specific miRNA sequences were deleted in
mouse models resulting in abnormal heart development in a large proportion of
the offspring. While these lethal effects were expected, other studies show a
more subtle role for miRNA in the heart. When miR-208 was eliminated, the mice
appeared normal. Only when their hearts were stressed, did defective responses
show up. These results show that comprehensive miRNA studies may be valuable in
the diagnosis of heart disease.
Neurological Diseases: Numerous reports have demonstrated the role of
miRNA in neural development. Evidence for a role in Parkinson's disease comes
from animal model studies published last year, showing that loss of miRNA may be
involved in the development and progression of the disease. In cell culture
experiments, transfer of small RNA fragments partially preserved miRNA-deficient
nerve cells. While these results and others point to an important role for miRNA
in neurodegenerative disorders, much more work is needed to delineate the exact
role of miRNA in this important area.
Immune Function Disorders: Recent miRNA deletion studies have revealed
a central role for them in the regulation of the immune response. The deletion
of miRNA-155 impaired T and B cell differentiation in germinal centers and
greatly decreased antibody and cytokine production. Two additional studies
deleting miRNA-181 and 223 were found to control T cell response and granulocyte
production, respectively. As more roles for miRNA in the immune response are
found, the list of immune function disorders with an miRNA component is certain
to expand.
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RT² miRNA PCR technology has
been developed to facilitate
accurate genome-wide miRNA
analysis with simple real-time
PCR assays. Read
more.
Detecting every miRNA across the entire genome in a specific and sensitive
way is a very challenging technology task. Many miRNA family members and
otherwise distinct miRNA species have very similar sequences. Moreover, other
RNA species such as snRNA, tRNA, mRNA, and rRNA can cause non-specific
amplification, making the specific analysis of mature miRNA even more
problematic. SABiosciences' proprietary miRNA technologies, including a miRNA
isolation technique, primer design methodology, and assay formulation for
universal reverse transcription specific to miRNA, address these challenges.
SABiosciences' complete miRNA PCR System includes:
- RT² miRNA Arrays and Assays
- RT² miRNA First Strand Kit
- RT² SYBR® Green PCR Master Mix
- RT² qPCR-Grade™ Small RNA Isolation Kit
RT² miRNA PCR Arrays and Assays dramatically improve the specificity for and
discrimination of closely related miRNA sequences and distinguishes mature miRNA
from other RNA species. Our miRNA PCR Arrays include build-in control elements
to insure the quality of your experimental data. The free data analysis software
takes your raw threshold cycle data and automatically generates figures and
tables ready for publication. With the RT² miRNA PCR Assay and Arrays, you can
expect:
- Sensitivity: As little as 0.5 µg total RNA needed
- Multi-Sequence Flexibility: Analyze one to 376 sequences
simultaneously
- Simplicity: As easy as a real-time PCR experiment
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