INTERFEROMICS

INDEX

TOPIC PAGE
No.

1) INTRODUCTION 3

2) RNA INTERFERENCE 5

3) DISCOVERY 6

4) CELLULAR MECHANISM
8

5) MicroRNA 9
6) RESEARCH ARTICLES 11

7) RNAI SYSTEM IS AN IN VITRO TRANSCRIPTION SYSTEM 11

8) HIGH-THROUGHPUT RNAI SCREENING IN CULTURED 11

CELLS: A USER’S GUIDE
9) CONCLUSION 14

10) REFERENCE

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INTRODUCTION:
Interferomics is a the study of biological events that take place post-transcriptomic pre-translatomically.
It defines one of many levels in an emerging field of Life sciences known as Systems Biology. Systems
biology is a term used to describe a number of trends in bioscience research, and a movement which
draws on those trends. Proponents describe systems biology as a biology-based inter-disciplinary study
field that focuses on complex interactions in biological systems, claiming that it uses a new perspective
(holism instead of reduction). Particularly from year 2000 onwards, the term is used widely in the
biosciences, and in a variety of contexts. An often stated ambition of systems biology is the modeling
and discovery of emergent properties, properties of a system whose theoretical description is only
possible using techniques which fall under the remit of systems biology.
According to the interpretation of Systems Biology as the ability to obtain, integrate and analyze
complex data from multiple experimental sources using interdisciplinary tools, some typical technology
platforms are:
Phenomics: Organismal variation in phenotype as it changes during its life span.
Genomics: Organismal deoxyribonucleic acid (DNA) sequence, including intra-organisamal cell
specific variation. (i.e. Telomere length variation etc.)
Epigenomics / Epigenetics: Organismal and corresponding cell specific transcriptomic regulating
factors not empirically coded in the genomic sequence. (i.e. DNA methylation, Histone Acetelation
etc.).
Transcriptomics: Organismal, tissue or whole cell gene expression measurements by DNA microarrays
or serial analysis of gene expression.
Interferomics: Organismal, tissue, or cell level transcript correcting factors (i.e. RNA interference)
Translatomics / Proteomics: Organismal, tissue, or cell level measurements of proteins and peptides
via two-dimensional gel electrophoresis, mass spectrometry or multi-dimensional protein identification
techniques (advanced HPLC systems coupled with mass spectrometry). Sub disciplines include
phosphoproteomics, glycoproteomics and other methods to detect chemically modified proteins.
Metabolomics: Organismal, tissue, or cell level measurements of all small-molecules known as
metabolites.
Glycomics: Organismal, tissue, or cell level measurements of carbohydrates.
Lipidomics: Organismal, tissue, or cell level measurements of lipids.
In addition to the identification and quantification of the above given molecules further techniques
analyze the dynamics and interactions within a cell. This includes:

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Interactomics: Organismal, tissue, or cell level study of interactions between molecules. Currently the
authoritative molecular discipline in this field of study is protein-protein interactions (PPI), although the
working definition does not pre-clude inclusion of other molecular disciplines such as those defined
here.
Fluxomics: Organismal, tissue, or cell level measurements of molecular dynamic changes over time.
Biomics: Systems analysis of the biome.
The investigations are frequently combined with large scale perturbation methods, including gene-based
(RNAi, mis-expression of wild type and mutant genes) and chemical approaches using small molecule
libraries. Robots and automated sensors enable such large-scale experimentation and data acquisition.
These technologies are still emerging and many face problems that the larger the quantity of data
produced, the lower the quality. A wide variety of quantitative scientists (computational biologists,
statisticians, mathematicians, computer scientists, engineers, and physicists) are working to improve the
quality of these approaches and to create, refine, and retest the models to accurately reflect observations.
The systems biology approach often involves the development of mechanistic models, such as the
reconstruction of dynamic systems from the quantitative properties of their elementary building blocks.
For instance, a cellular network can be modelled mathematically using methods coming from chemical
kinetics and control theory. Due to the large number of parameters, variables and constraints in cellular
networks, numerical and computational techniques are often used.
Other aspects of computer science and informatics are also used in systems biology. These include:
New forms of computational model, such as the use of process calculi to model biological processes
(notable approaches include stochastic π-calculus, BioAmbients, Beta Binders, BioPEPA and Brane
calculus) and constraint-based modeling. Integration of information from the literature, using
techniques of information extraction and text mining.
Development of online databases and repositories for sharing data and models, approaches to database
integration and software interoperability via loose coupling of software, websites and databases, or
commercial suits.
Development of syntactically and semantically sound ways of representing biological models.
RNA Interference is one such cellular mechanism that falls in this category and is to-date the only
known inteferomic phenomenon. Drs Andrew Z. Fire and Craig C. Mello were awarded the Nobel Prize
in Physiology or Medicine in the year of 2006 for their collective discovery of RNA interference.
As a field of study, particularly, the study of the interactions between the components of biological
systems, and how these interactions give rise to the function and behavior of that system (for example,
the enzymes and metabolites in a metabolic pathway).

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As a paradigm, usually defined in antithesis to the so-called reductionist paradigm (biological
organisation), although fully consistent with the scientific method. The distinction between the two
paradigms is referred to in these quotations:

"The reductionist approach has successfully identified most of the components and many of the
interactions but, unfortunately, offers no convincing concepts or methods to understand how system
properties emerge...the pluralism of causes and effects in biological networks is better addressed by
observing, through quantitative measures, multiple components simultaneously and by rigorous data
integration with mathematical models" Science

"Systems biology is about putting together rather than taking apart, integration rather than reduction. It
requires that we develop ways of thinking about integration that are as rigorous as our reductionist
programmes, but different. It means changing our philosophy, in the full sense of the term" Denis Noble

As a series of operational protocols used for performing research, namely a cycle composed of theory,
analytic or computational modelling to propose specific testable hypotheses about a biological system,
experimental validation, and then using the newly acquired quantitative description of cells or cell
processes to refine the computational model or theory. Since the objective is a model of the interactions
in a system, the experimental techniques that most suit systems biology are those that are system-wide
and attempt to be as complete as possible. Therefore, transcriptomics, metabolomics, proteomics and
high-throughput techniques are used to collect quantitative data for the construction and validation of
models.
As the application of dynamical systems theory to molecular biology.
As a socioscientific phenomenon defined by the strategy of pursuing integration of complex data about
the interactions in biological systems from diverse experimental sources using interdisciplinary tools
and personnel.
This variety of viewpoints is illustrative of the fact that systems biology refers to a cluster of
peripherally overlapping concepts rather than a single well-delineated field. However the term has
widespread currency and popularity as of 2007, with chairs and institutes of systems biology
proliferating worldwide.
RNA INTERFERENCE:
RNA interference (RNAi) is a system within living cells that takes part in controlling which genes are
active and how active they are. Two types of small RNA molecules – microRNA (miRNA) and small
interfering RNA (siRNA) – are central to RNA interference. RNAs are the direct products of genes, and
these small RNAs can bind to other specific RNAs (mRNA) and either increase or decrease their

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activity, for example by preventing a messenger RNA from producing a protein. RNA interference has
an important role in defending cells against parasitic genes – viruses and transposons – but also in
directing development as well as gene expression in general.
DISCOVERY:
The discovery of RNAi was preceded first by observations of transcriptional inhibition by antisense
RNA expressed in transgenic plants, and more directly by reports of unexpected outcomes in
experiments performed by plant scientists in the United States and the Netherlands in the early 1990s. In
an attempt to alter flower colors in petunias, researchers introduced additional copies of a gene encoding
chalcone synthase, a key enzyme for flower pigmentation into petunia plants of normally pink or violet
flower color. The overexpressed gene was expected to result in darker flowers, but instead produced less
pigmented, fully or partially white flowers, indicating that the activity of chalcone synthase had been
substantially decreased; in fact, both the endogenous genes and the transgenes were downregulated in
the white flowers. Soon after, a related event termed quelling was noted in the fungus Neurospora
crassa, although it was not immediately recognized as related. Further investigation of the phenomenon
in plants indicated that the downregulation was due to post-transcriptional inhibition of gene expression
via an increased rate of mRNA degradation. This phenomenon was called co-suppression of gene
expression, but the molecular mechanism remained unknown.
Not long after, plant virologists working on improving plant resistance to viral diseases observed a
similar unexpected phenomenon. While it was known that plants expressing virus-specific proteins
showed enhanced tolerance or resistance to viral infection, it was not expected that plants carrying only
short, non-coding regions of viral RNA sequences would show similar levels of protection. Researchers
believed that viral RNA produced by transgenes could also inhibit viral replication.[143] The reverse
experiment, in which short sequences of plant genes were introduced into viruses, showed that the
targeted gene was suppressed in an infected plant. This phenomenon was labeled "virus-induced gene
silencing" (VIGS), and the set of such phenomena were collectively called post transcriptional gene
silencing.
After these initial observations in plants, many laboratories around the world searched for the
occurrence of this phenomenon in other organisms. Craig C. Mello and Andrew Fire's 1998 Nature
paper reported a potent gene silencing effect after injecting double stranded RNA into C. elegans. In
investigating the regulation of muscle protein production, they observed that neither mRNA nor
antisense RNA injections had an effect on protein production, but double-stranded RNA successfully
silenced the targeted gene. As a result of this work, they coined the term RNAi. Fire and Mello's
discovery was particularly notable because it represented the first identification of the causative agent

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for the phenomenon. Fire and Mello were awarded the Nobel Prize in Physiology or Medicine in 2006
for their work.

The RNAi pathway is found in many eukaryotes including animals and is initiated by the enzyme Dicer,
which cleaves long double-stranded RNA (dsRNA) molecules into short fragments of ~20 nucleotides.
The siRNA will be unwound into two ssRNA, namely the passenger strand and the guide strand. The
passenger strand will be degraded, and the guide strand is incorporated into the RNA-induced silencing
complex (RISC). The most well-studied outcome is post-transcriptional gene silencing, which occurs
when the guide strand base pairs with a complementary sequence of a messenger RNA molecule and
induces cleavage by Argonaute, the catalytic component of the RISC complex. This process is known to
spread systemically throughout the organism despite initially limited molar concentrations of siRNA.
The selective and robust effect of RNAi on gene expression makes it a valuable research tool, both in
cell culture and in living organisms because synthetic dsRNA introduced into cells can induce
suppression of specific genes of interest. RNAi may also be used for large-scale screens that
systematically shut down each gene in the cell, which can help identify the components necessary for a
particular cellular process or an event such as cell division. Exploitation of the pathway is also a
promising tool in biotechnology and medicine.
Historically, RNA interference was known by other names, including cosuppression, post transcriptional
gene silencing, and quelling. Only after these apparently unrelated processes were fully understood did
it become clear that they all described the RNAi phenomenon. In 2006, Andrew Fire and Craig C. Mello

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shared the Nobel Prize in Physiology or Medicine for their work on RNA interference in the nematode
worm C. elegans, which they published in 1998.
CELLULAR MECHANISM:
The dicer protein from Giardia intestinalis, which catalyzes the cleavage of dsRNA to siRNAs. The
RNase domains are colored green, the PAZ domain yellow, the platform domain red, and the connector
helix blue.RNAi is an RNA-dependent gene silencing process that is controlled by the RNA-induced
silencing complex (RISC) and is initiated by short double-stranded RNA molecules in a cell's
cytoplasm, where they interact with the catalytic RISC component argonaute. When the dsRNA is
exogenous (coming from infection by a virus with an RNA genome or laboratory manipulations), the
RNA is imported directly into the cytoplasm and cleaved to short fragments by the enzyme Dicer. The
initiating dsRNA can also be endogenous (originating in the cell), as in pre-microRNAs expressed from
RNA-coding genes in the genome. The primary transcripts from such genes are first processed to form
the characteristic stem-loop structure of pre-miRNA in the nucleus, then exported to the cytoplasm to be
cleaved by Dicer. Thus, the two dsRNA pathways, exogenous and endogenous, converge at the RISC
complex.
dsRNA cleavageEndogenous dsRNA initiates RNAi by activating the ribonuclease protein Dicer which
binds and cleaves double-stranded RNAs (dsRNAs) to produce double-stranded fragments of 20–25
base pairs with a 2 nucleotide overhang at the 3' end. Bioinformatics studies on the genomes of multiple
organisms suggest this length maximizes target-gene specificity and minimizes non-specific effects.
These short double-stranded fragments are called small interfering RNAs (siRNAs). These siRNAs are
then separated into single strands and integrated into an active RISC complex. After integration into the
RISC, siRNAs base-pair to their target mRNA and induce cleavage of the mRNA, thereby preventing it
from being used as a translation template.
Exogenous dsRNA is detected and bound by an effector protein, known as RDE-4 in C. elegans and
R2D2 in Drosophila, that stimulates dicer activity.This protein only binds long dsRNAs, but the
mechanism producing this length specificity is unknown.These RNA-binding proteins then facilitate
transfer of cleaved siRNAs to the RISC complex.
In C. elegans, this initiation response is amplified by the cell by the synthesis of a population of
'secondary' siRNAs using the dicer-produced initiating or 'primary' siRNAs as templates. These siRNAs
are structurally distinct from dicer-produced siRNAs and appear to be produced by an RNA-dependent
RNA polymerase (RdRP).

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MicroRNA
The stem-loop secondary structure of a pre-microRNA from Brassica oleracea.Main article: MicroRNA
MicroRNAs (miRNAs) are genomically encoded non-coding RNAs that help regulate gene expression,
particularly during development. The phenomenon of RNA interference, broadly defined, includes the
endogenously induced gene silencing effects of miRNAs as well as silencing triggered by foreign
dsRNA. Mature miRNAs are structurally similar to siRNAs produced from exogenous dsRNA, but
before reaching maturity, miRNAs must first undergo extensive post-transcriptional modification. An
miRNA is expressed from a much longer RNA-coding gene as a primary transcript known as a pri-
miRNA which is processed, in the cell nucleus, to a 70-nucleotide stem-loop structure called a pre-
miRNA by the microprocessor complex. This complex consists of an RNase III enzyme called Drosha
and a dsRNA-binding protein Pasha. The dsRNA portion of this pre-miRNA is bound and cleaved by
Dicer to produce the mature miRNA molecule that can be integrated into the RISC complex; thus,
miRNA and siRNA share the same cellular machinery downstream of their initial processing.

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The siRNAs derived from long dsRNA precursors differ from miRNAs in that miRNAs, especially
those in animals, typically have incomplete base pairing to a target and inhibit the translation of many
different mRNAs with similar sequences. In contrast, siRNAs typically base-pair perfectly and induce
mRNA cleavage only in a single, specific target. In Drosophila and C. elegans, miRNA and siRNA are
processed by distinct argonaute proteins and dicer enzymes.
RISC activation and catalysisThe active components of an RNA-induced silencing complex (RISC) are
endonucleases called argonaute proteins, which cleave the target mRNA strand complementary to their
bound siRNA. As the fragments produced by dicer are double-stranded, they could each in theory
produce a functional siRNA. However, only one of the two strands, which is known as the guide strand,
binds the argonaute protein and directs gene silencing. The other anti-guide strand or passenger strand is
degraded during RISC activation. Although it was first believed that an ATP-dependent helicase
separated these two strands, the process is actually ATP-independent and performed directly by the
protein components of RISC The strand selected as the guide tends to be the one whose 5' end is least
paired to its complement, but strand selection is unaffected by the direction in which dicer cleaves the
dsRNA before RISC incorporation. Instead, the R2D2 protein may serve as the differentiating factor by
binding the more-stable 5' end of the passenger strand.
The structural basis for binding of RNA to the argonaute protein was examined by X-ray
crystallography of the binding domain of an RNA-bound argonaute protein. Here, the phosphorylated 5'
end of the RNA strand enters a conserved basic surface pocket and makes contacts through a divalent
cation (an atom with two positive charges) such as magnesium and by aromatic stacking (a process that
allows more than one atom to share an electron by passing it back and forth) between the 5' nucleotide
in the siRNA and a conserved tyrosine residue. This site is thought to form a nucleation site for the
binding of the siRNA to its mRNA target.
It is not understood how the activated RISC complex locates complementary mRNAs within the cell.
Although the cleavage process has been proposed to be linked to translation, translation of the mRNA
target is not essential for RNAi-mediated degradation. Indeed, RNAi may be more effective against
mRNA targets that are not translated. Argonaute proteins, the catalytic components of RISC, are
localized to specific regions in the cytoplasm called P-bodies (also cytoplasmic bodies or GW bodies),
which are regions with high rates of mRNA decay,miRNA activity is also clustered in P-bodies.
Disruption of P-bodies decreases the efficiency of RNA interference, suggesting that they are the site of
a critical step in the RNAi process.
Transcriptional silencingComponents of the RNA interference pathway are also used in many
eukaryotes in the maintenance of the organization and structure of their genomes. Modification of
histones and associated induction of heterochromatin formation serves to downregulate genes pre-

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transcriptionally; this process is referred to as RNA-induced transcriptional silencing (RITS), and is
carried out by a complex of proteins called the RITS complex. In fission yeast this complex contains
argonaute, a chromodomain protein Chp1, and a protein called Tas3 of unknown function.As a
consequence, the induction and spread of heterochromatic regions requires the argonaute and RdRP
proteins. Indeed, deletion of these genes in the fission yeast S. pombe disrupts histone methylation and
centromere formation, causing slow or stalled anaphase during cell division.In some cases, similar
processes associated with histone modification have been observed to transcriptionally upregulate genes.
The mechanism by which the RITS complex induces heterochromatin formation and organization is not
well understood, and most studies have focused on the mating-type region in fission yeast, which may
not be representative of activities in other genomic regions or organisms. In maintenance of existing
heterochromatin regions, RITS forms a complex with siRNAs complementary to the local genes and
stably binds local methylated histones, acting co-transcriptionally to degrade any nascent pre-mRNA
transcripts that are initiated by RNA polymerase. The formation of such a heterochromatin region,
though not its maintenance, is dicer-dependent, presumably because dicer is required to generate the
initial complement of siRNAs that target subsequent transcripts. Heterochromatin maintenance has been
suggested to function as a self-reinforcing feedback loop, as new siRNAs are formed from the
occasional nascent transcripts by RdRP for incorporation into local RITS complexes. The relevance of
observations from fission yeast mating-type regions and centromeres to mammals is not clear, as
heterochromatin maintenance in mammalian cells may be independent of the components of the RNAi
pathway.
Crosstalk with RNA editing:
The type of RNA editing that is most prevalent in higher eukaryotes converts adenosine nucleotides into
inosine in dsRNAs via the enzyme adenosine deaminase (ADAR). It was originally proposed in 2000
that the RNAi and A→I RNA editing pathways might compete for a common dsRNA substrate.Indeed,
some pre-miRNAs do undergo A→I RNA editing, and this mechanism may regulate the processing and
expression of mature miRNAs. Furthermore, at least one mammalian ADAR can sequester siRNAs
from RNAi pathway components. Further support for this model comes from studies on ADAR-null C.
elegans strains indicating that A→I RNA editing may counteract RNAi silencing of endogenous genes
and transgenes.

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RESEARCH ARTICLES:
1. RNAI SYSTEM IS AN IN VITRO TRANSCRIPTION SYSTEM:
T7 RiboMAX™ Express RNAi SystemThe T7 RiboMAX™ Express RNAi System is an in vitro
transcription system designed for producing milligram amounts of double-stranded RNA (dsRNA) in a
short amount of time. The dsRNA is free of protein and other contaminants and is suitable for use in
RNA interference (RNAi) in both mammalian and nonmammalian systems.
The T7 RiboMAX™ Express RNAi System can be used to synthesize short interfering RNAs (siRNAs)
of 21bp for use in mammalian systems. siRNAs synthesized in vitro have been demonstrated to be as
effective as chemically synthesized siRNAs for inducing RNAi in mammalian cells.
In addition, the T7 RiboMAX™ Express RNAi System can be used for the synthesis of dsRNA
molecules of approximately 200bp or greater, which can be applied to nonmammalian systems. Two
complementary RNA strands are synthesized from DNA template (either plasmid or PCR product). The
resulting RNA strands are annealed after the transcription reaction to form dsRNA. Any remaining
single-stranded RNA and DNA template are removed with a nuclease digestion step. The dsRNA is then
purified by isopropanol precipitation and can be introduced into the organism of choice for RNAi
applications.
2. HIGH-THROUGHPUT RNAI SCREENING IN CULTURED CELLS: A USER’S GUIDE
RNA interference has re-energized the field of functional genomics by enabling genome-scale loss-of-
function screens in cultured cells. Looking back on the lessons that have been learned from the first
wave of technology developments and applications in this exciting field, we provide both a user’s guide
for newcomers to the field and a detailed examination of some more complex issues, particularly
concerning optimization and quality control, for more advanced users. From a discussion of cell lines,
screening paradigms, reagent types and read-out methodologies, we explore in particular the
complexities of designing optimal controls and normalization strategies for these challenging but
extremely powerful studies.
RNA interference: the new somatic cell genetics?
RNAi is evolving into a powerful tool for manipulating gene expression in mammalian cells with
potential utility for investigating gene function, for high-throughput, function-based genetic screens and
potentially for development as a therapeutic tool.
RNAi in mammals:
Given the strong conservation of RNAi-related genes in vertebrates, including Dicer and Argonaute
family members, the expectation was that RNAi would operate in mammalian cells in some capacity.
The first glimpse of RNAi in mammals came from injections of long dsRNAs (?500 nt, similar to those

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used to trigger RNAi in invertebrate systems) into mouse embryos, which resulted in sequence-specific
gene silencing. Several groups,including our own, extended these findings to embryonal cell Lines.
Biochemical and genetic evidence from these studies suggested that RNAi operates in at least a subset
of mammalian cell types, in a Dicer-dependent manner via posttranscriptional mechanisms In somatic
cells, however, the use of conventional dsRNA triggers is limited by antiviral/interferon responses,
including the PKR and RNaseL pathways, which trigger generalized translational repression and
apoptosis in response to dsRNA of >30 bp in length. Even where PKR activity is removed from somatic
cells, by either viral inhibitors or targeted disruption, long dsRNA still triggers a residual nonspecific
repression of gene expression.One way around these nonspecific dsRNA responses is to simply create
dsRNA triggers of <30 bp in length. In the past year, two short dsRNA structures have emerged, which
evoke sequence specific gene silencing in somatic cells without activating antiviral responses. These are
the small interfering RNAs (siRNAs) and the short hairpin RNAs (shRNAs). Both are modeled after
biologically active structures in the RNAi pathway:
Dicer cleavage products and small temporal RNAs, respectively.Tuschl and colleagues and Caplen and
colleagues (Elbashir et al., 2001; Caplen et al., 2001) first demonstrated that small dsRNAs, resembling
siRNAs from other systems, induce sequence-specific gene silencing when transiently transfected into
mammalian cells.These small interfering RNAs (siRNAs) are chemically synthesized emulations of
Dicer cleavage products, which are short RNA duplexes ?19 nt in length with 2 nt 3 overhangson each
strand. The siRNAs presumably bypass the requirement for Dicer and enter the silencing pathway by
incorporation into RISC complexes. The use of siRNAs has been recently reviewed, in detail, and
resources for the design and use of siRNAs are available from Tom Tuschl’s laboratory online
(http://www.mpibpc.gwdg.de/abteilungen/100/105/sirna.html). As an alternative strategy, we and
others have developed in vivo expression constructs for small dsRNA triggers in mammalian cells,
which resemble endogenously expressed hairpin RNAs (Paddison et al., 2002b; Brummelkamp et al.,
2002; Paul
et al., 2002; Sui et al., 2002;Yu et al., 2002; Zeng et al., 2002). This approach uses small inverted
repeats (19–29 nt) expressed from RNA polymerase III promoter to create short hairpin RNAs
(shRNAs), which can then be processed by Dicer and shunted into the RNAi pathway (Figure 1).
However, siRNAs can also be produced in vivo by the expression of complementary 19 or 21 nt RNAs
from separate RNA polymerase III transcription units (Lee et al., 2002; Miyagishi and Taira, 2002; Yu
et al., 2002). For some studies, expressed dsRNA triggers have potential advantages over siRNAs when
combined with well-worn strategies for stable and inducible gene expression in vitro and in vivo. The
details of these strategies are further discussed below. One of the major differences between mammalian
cell RNAi and the response observed, for example, in C. elegans is the apparent lack of amplification of

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the RNAi effect or of “transitive RNAi” (Sijen et al., 2001). In C. elegans, “amplification” may
contribute to heritable, systemic gene silencing . According to one model, amplification of the dsRNA
signal is initially mediated by RNA-dependent RNA polymerases (RdRP). An RNA degradation product
(e.g., an siRNA) may prime RdRPs along the mRNA template, resulting in the production of dsRNA
homologous to sequences 5 (i.e., upstream) of the initially targeted sequence (Sijen et al., 2001). When
combined with transport, amplification results in a self-propagating silencing effect throughout the
organism. In mammalian cell systems, however, transient transfection of RNAi triggers, e.g., long
dsRNA, siRNAs, or shRNAs, results in a transient effect, lasting 2–7 days. The longevity of silencing is
likely dependent on gene expression homeostasis (e.g., abundance of mRNA and protein, stability of the
protein, transcriptional feedback loops, etc.), the half-life of the silencing complex itself, and cell
division, which serves to dilute the effect over time.
CONCLUSION:
The field of life sciences has grown beyond leaps and bounds and since the advent of bioinformatics
many changes have taken place and led to a new dimension and view for the branch. Systems biology in
particular has revolutionized and laid much bigger platform to unravel the secrets of biology. It is a
branch which can describe new trends in bioscience research and helps in understanding the movement
of such trends.
Since 2000 the term systems biology is in the circle of biosciences and has been used in large context
and often used in the area of modeling, discovery of some properties. In technical terms it means the
examination of structure and dynamics of various functions at cellular and organismal level instead of
understanding the characteristics of isolated parts of cells.

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REFERENCE :

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