ANTISENSE RNA TECHNOLOGY
FLAVR SAVR TOMATO
GENE SILENCING
RNA INTERFERENCE
miRNA
ANTISENSE RNA
TECHNOLOGY
In any gene the DNA strand which is transcribed. This
strand is called antisense strand
The DNA strand that is not transcribed is called sense
strand as the mRNA formed has a similar sequence as
that of the sense strand
The mRNA formed is called sense mRNA
An antisense gene is produced by iverting the
orientation of the protein encoding region in relation to
its promoter. As a result the natural sense strand of
the gene becomes oriented in the 3’-5’ direction with
respect to its promoter and is transcribed
The RNA produced by the inverted strand is called
antisense RNA
When an antisense gene is present in the same
nucleus along with the normal gene, transcription
of the two genes yield antisense and sense RNA
Since the sense and the antisense RNA are
complementary to each other, they could pair to
produce dsRNA molecule
The dsRNA is thus not available for translation
The dsRNA is attacked and degraded by dsRNA
specific RNAase. Hence the gene expression is
suppressed and induces its cleavage by catalytic
part of a RISC complex called argonaute.
GENE SILENCING
Frequent expression of transgene is suppressed in the
subsequent generations of transgenic plants. This is
called gene silencing
Gene silencing is often acompanied by DNA methylation
of transgenes. In general higher the no. of copies of a
transgene, more frequent is their hyper methylation and
transgene inactivation
Gene silencing mechanisms are of two types
1. transcriptional silencing (no mRNA is formed)
2. Post transcriptional silencing (no proteing is formed)
Transcriptional silencing is generally due to promoter
methylation, while post transcriptional silencing can
involve methylation of the coding sequence
RNA INTERFERENCE
It is a biological process in which RNA
molecules inhibit gene expression
Silencing of homologous gene expression
triggered by dsRNA is called RNAi
Introduction of long dsRNA into cells of
plants invertebrate and mammals lead to a
sequence specific degradation of the
homologous gene transcripts
The long dsRNA molecule are cleaved by an
RNAase III enzyme called Dicer
This generates small 21-23 nucleotide long dsA molecule
called small interfering RNA (siRNA)
The small interfering RNA molecule binds to a protein
complex called RNA induced silencing complex (RISC). This
complex contains a helicase activity that unbinds the two
strands of RNA molecules
The antisense RNA strands so generated pair with the
target RNA molecule and an endonuclease activity then
hydrolyses the target RNA at the site where the antisense
strand is bound
RNAi serves as an antiviral mechanism (defence)
RNAi is now becoming a powerful tool and is widely used for
the analysis of the gene functions in invertebrates, plant
and mammals.
RNA Interference (RNAi).MP4
RNA interference animation with Malayalam subtitles.flv
miRNA
microRNA is a short single stranded RNA molecule
that plays an important role in gene regulation
miRNA is single stranded and attaches to the
specific regions of the mRNA thus inhibiting
translation and protein synthesis (untranslated
region- UTR)
miRNA are found in eukaryotic cells that bind to
the untranslated region at the 3’ end of mRNA
sequence
miRNA’s have all sorts of functions in physiology
from cell differention, proliferation, apoptosis etc.
miRNAs are widely used in therapeutics now
FLAVR –SAVR TOMATO
An application of antisense RNA technology
is in the production of a slow ripening
tomato called FLAVR SAVR tomato
These tomato have increased shelf life as
they ripen slowly
This is achieved by suppression of
polygalacturonase producing genes by
antisense RNA technology
Polygalactouronase is the pectin degrading
enzyme which softens the fruits
