RNA interference (RNAi): Cellular process by which an mRNA is targeted for degradation by a dsRNA with a strand complementary to a fragment of such mRNA.
RNA interference (RNAi): Cellular process by which an mRNA is targeted for degradation by a dsRNA with a strand complementary to a fragment of such mRNA.
Introduction
Transcriptome analysis
Goal of functional genomics
Why we need functional genomics
Technique
1. At DNA level
2.At RNA level
3. At protein level
4. loss of function
5. functional genomic and bioinformatics
Application
Latest research and reviews
Websites of functional genomics
Conclusions
Reference
RNAi is a powerful, conserved biological process through which the small, double-stranded RNAs specifically silence the expression of homologous genes, largely through degradation of their cognate mRNA.
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
Introduction
Transcriptome analysis
Goal of functional genomics
Why we need functional genomics
Technique
1. At DNA level
2.At RNA level
3. At protein level
4. loss of function
5. functional genomic and bioinformatics
Application
Latest research and reviews
Websites of functional genomics
Conclusions
Reference
RNAi is a powerful, conserved biological process through which the small, double-stranded RNAs specifically silence the expression of homologous genes, largely through degradation of their cognate mRNA.
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
Antisense RNA is the complementary RNA to the protein-coding messenger mRNA. The antisense mRNA binds to sense mRNA and the translation was restricted, that is called translation arrest. The nature antisense RNA technology present in hok &sok in E.coli R1 plasmid. The artificial or purpose antisense RNA technology best example is flavr savr tomato.
Antisense RNA Technology for crop improvement.pptxSanyamPatel2
Improvement of crops by following genetic engineering techniques:
Gene silencing
Post transcription silencing
Post translation silencing
Antisense RNA
RNA interference
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
3. INTRODUCTION
Antisense RNA is a single-stranded RNA that is
complementary to a messenger RNA (mRNA) strand
transcribed within a cell
Antisense RNA introduced into a cell to inhibit
translation of a complementary mRNA by base
pairing to it and creating barrier to the translation
machinery.
E.g.
hok/sok system of the E. coli R1 plasmid.
3
4.
This translational arrest causes reduced amount of
protein expression.
Well-known examples of GM plants produced by
this technology-
The Flavr Savr tomato ,
Two cultivars of ring spot-resistant papaya.
4
After 45 days….
6. Diff. between antisense
technology & RNAi
The intended effect in both will be same i.e. gene
silencing but the processing is little but different.
Antisense technology degrades RNA by enzymes
RNaseH while RNAi employed the enzyme DICER
to degrade the m RNA.
RNAi are twice larger than the antisense
oligonucleotide.
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7. HISTORY
First time at “ free university of Amsterdam”, used
antisense RNA technology against the gene
determining flower color of petunia .
Antisense effect first demonstrated by zemencnick &
Stephenson in 1970 on “Rous sarcoma virus”.
First time antisense oligonucleotides are synthesized
by Eckstein and colleagues.
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8.
In 1995 Guo and Kemp hues:
injection of either antisense or sense RNAs in the
germ line of C. elegans was equally effective at
silencing homologous target genes.
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9. Nature’s antisense system
There is HOK(host killing)/SOK(suppress killing) system
in R1 plasmid in E.Coli.
when E. coli cell undergoes division , daughter cell inherit
hok gene & sok gene from parent. But due to short life of
cell, the sok gene is get degraded. So in normal cell, hok
gene get over expressed & cell get die.
But when R1 plasmid is get inherited , it having the sok
gene & sok promoter.
Then it transcripts sok gene & it is get overexpressed
against hok gene.
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11. MECHANISM
In this technique, Short segments of single stranded
RNA are introduced.
These oligonucleotides are complementary to the
mRNA, which physically bind to the mRNA.
So , they block the expression of particular gene.
In case of viruses, antisense oligonucleotides inhibit
viral replication with blocking expression of
integrated proviral genes.
Usually consist of 15–20 nucleotides.
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12.
Translation of mRNA may be blocked by two
possible mechanisms , These are:-
1] by base specific hybridization – which prevents
access by translation machinery i.e. “hybridization
arrest”.
2] by forming RNA/DNA duplex which is
recognized by nuclease RNaseH , specific for digesting
RNA in an RNA/DNA duplex.
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13.
RNaseH is a non-specific endonuclease, catalyzes the
cleavage of RNA via hydrolytic mechanism.
RNaseH has ribonuclease activity cleaves the 3’-O-P
bond of RNA in a DNA/RNA duplex.
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14.
Unique DNA sequence
Efficient cellular uptake
Minimal nonspecific binding
Target specific hybridization
Non-toxic antisense construct
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Characteristics of antisense
oligonucleotides
15.
The antisense technology can be modified in THREE
modes because of chemical modifications of the
oligonucleotides.
These modes are due to activation of RNaseH &
internucleotides linkages which do not activate
enzyme.
15
Approaches
16.
The antisense oligonucleotides binds the target
sequence causing both “hybridisation arrest ” &
“RNaseH activation”.
Degradation of mRNA by RNaseH results into
release of oligonucleotides.
They may bind to other copies of target mRNA.
These oligonucleotides are also susceptible to other
nucleases.
This a major parameter affecting catalytic mode of
degradation.
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1st approach
17.
In this, antisense oligonucleotides binds to target
sequence result in translation arrest but they do not
activate enzyme RNaseH.
Oligoribonucleotides & there analogues ,
oligodeoxyribonucleotides , various non phosphate
& phosphate internucleotides linkages fall in this
category.
They show resistance against nucleuses enzyme and
never get degraded by them.
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2nd approach
18.
They also show effective translational arrest .
But the major problem is that they are generally
required higher molar concentrations than those
which activate RNaseH.
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19.
It combines features of both previous approaches.
They contains both internucleotides linkages which
are responsible for RNaseH activation & which
shows resistance against them.
Digestion of mRNA target in RNA-DNA duplex
releases oligonucleotides which are resistance
against nuclease enzyme, hence are more effective
than oligonucleotides in 1st approach.
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3rd approach
20.
They may form hybrids of
oligodeoxyribonucleotides & Oligoribonucleotides.
The antiviral activity of an antisense oligonucleotides
depends usually on specific binding to a target
nucleic acid.
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22.
Thomas and coworkers coined the term ‘ribozymes’.
These are RNA molecules which have catalytic
activity which degrade nucleotides .
Ribozyme Bind to the target RNA moiety and
inactivate it by cleaving the phosphodiester
backbone at a specific cutting site.
Ribozyme destroy RNA that carries the massage of
disease.
These are effectively used against HIV virus.
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Ribozymes
24. APPLICATION
1. Flavr Savr tomato-antisense
RNA used against an enzyme
polygalacturonase, an softening enzyme which is responsible
for ripening.
2. Transgenic ACMV-resistant cassava plants* –
Used against African cassava mosaic virus
(ACMV) which causes cassava mosaic disease causing major
economic loss in Africa.
3. Formivirsen-is
the first antiviral drug developed against CMV.
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26. conclusion
Antisense technology shows potential for diverse
application to field of basic research & therapy.
One of the most approved approaches for inactivating a
single specific gene.
But it may sometime give undesirable effect.
Generally , antisense RNA still lack effective design,
biological activity, and efficient route of administration.
Antisense technologies form a very powerful weapon for
studying gene function and for discovering more specific
treatments of disease.
26
27.
Attempts are made to genetically engineer transgenic
plants to express antisense RNA instead activate the
RNAi pathway, although the processes result in
“gene silencing”.
27
28. References :-
A textbook of biotechnology 2nd edition by H. D.
Kumar
www.youtube.com
Nature biotechnology.
www.ncbi.nlm.nih.com (PubMed ID 17173627)*
www.google.com
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