CURRENT UPDATES

ON GM CROPS
PRESENTED TO: DR. SABA IRSHAD KHOKHAR
PRESENTED BY: ADEEL QASIM
ROLL NUMBER : 06
CURRENT UPDATES ON GM CROPS

• Current studies regarding GM Crops

• Current genetic modification techniques for GMO’s
identification and gene editing
• Current frameworks for GMO’s implementation
CURRENT STUDIES REGARDING GM CROPS

• Mitochondrial pyruvate carrier 1 mediates abscisic acid-

regulated stomatal closure and the drought response by
affecting cellular pyruvate content in Arabidopsis
thaliana
 ABSCISIC ACID AND TREATMENT

PLANT

But mPC1 mutant accumulates Pyruvate content increases and

the pyruvate in cytosol thereby ROS produced in guard cells
encouraging ROS production and
accelerating stomatal closure.
 INCREASE IN PYRUVATE CONTENT

• Activate slow type anion channels which results in the

efflux of anions, which in turn reduces the turgor of guard
cells to close the stomata.
 FUNCTIONAL MPC1

• Also has role in this process, it allows pyruvate to move

towards inner mitochondrial membrane instead of
remaining in cytosol, due to decrease in pyruvate content,
stomatal closure does not happen.
PROPOSED MODEL FOR THE ROLE OF ATMPC1 IN THE
REGULATION OF STOMATAL MOVEMENT AND THE
DROUGHT RESPONSE
CURRENT STUDIES REGARDING GM CROPS

Plant molecular farming (PMF), defined as the practice of using plants to produce
human therapeutic proteins, has received worldwide interest.
• Identifying potential genes suitable for PMF and general approaches is becoming more simple
and straight forward. Facilitated by the rapid progress in genomics, proteomics, and
bioinformatics, a greater number of useful genes are being identified and characterized.
Additionally, relatively routine molecular methods have become available for placing the genes
of interest into plant expression vectors and transforming them into plants.
DIAGRAMMATIC ILLUSTRATION OF PMF:
EXAMPLES OF PLANT-DERIVED PHARMACEUTICALS:
COMPARISON OF DIFFERENT EXPRESSION PLATFORMS
FOR THE PRODUCTION OF PHARMACEUTICALS:
ZOONOSIS:

• Zoonosis can be defined as transmission of disease between

human and animals that happens due to interaction between
these two populations. Zoonosis not only interrupts human
health but it also affects wild life and livestock industry.
Recently, more than 65% of emerging infectious diseases in
humans have been reported to originate from zoonotic
pathogens.
 CURRENT GENETIC MODIFICATION TECHNIQUES FOR
GMO IDENTIFICATION AND GENE EDITING:

whole genome shotgun sequencing:

In whole genome shotgun sequencing the entire genome is broken up into small fragments of DNA. These
fragments are often of varying sizes, ranging from 2-20 kilobases (2,000-20,000 base pairs) to 200-300 kilobases
(200,000-300,000 base pairs). These fragments are sequenced to determine the order of the DNA bases, A, C,
G and T. The sequenced fragments are then assembled together by computer programs that find where
fragments overlap.
• Approaches to detect, characterize, and identify GMOs by application of whole genome
shotgun sequencing. First the DNA is extracted from the sample and purified
• (A). Longer fragments are optionally split into shorter fragments to obtain a sequencing library with
fragments of a desired size range; for example, approximately 500 bp for paired-end sequencing or 2–10 kbp
for mate-pair sequencing
• Continued….
• (B) Capture enrichment from the sequencing library can optionally be performed before
sequencing.
• (C) Raw sequencing reads are quality filtered before bioinformatics analysis.
• (D) Three types of available sequence databases can be available for GMO analyses
• (E) The full sequence of the taxon (reference genome), the full sequence of the insertion vector
used to create the GMO, and a collection (more or less complete) of sequence elements/motifs
associated with various GMOs (GMO sequence elements database).
• Mapping of the reads to these databases can result in identification of F perfect matches (i.e.,
concordant mapping), G reads with matching and orphan mates (i.e., discordant mapping), H
nonmapping (i.e., unmapped reads), and I chimeric reads mapping partially to one database
sequence and partially to another database sequence (i.e., split reads).
• The illustrated orphan reads in G and I sometimes map to other sequences in the same or other
databases, and in such cases provide useful information for further mapping. Notably, some
sequencing technologies produce single reads, not paired/mated reads.
• In these cases only F, H, and can be observed. Mapping can be done against only one of the
references or against two or all databases. Depending on the order and results of the mapping, the
outcome is usually a subset of sequence reads that can be used to infer the sequence of the
genetically modified insert and its insertion locus.
• Perfectly mapped reads confirm the presence of a particular sequence motif (J). Paired/mated
reads can facilitate the assembly of reads into contigs (K).
• Single reads can be assembled to shorter contigs (L and M) that can successively be assembled
into longer contigs (N).
CRISPER CAS9:

• The Clustered Regularly Interspaced Short Palindromic Repeats associated Cas9/sgRNA

system is a novel targeted genome-editing technique derived from bacterial immune
system. It is an inexpensive, easy, most user friendly and rapidly adopted genome editing
tool transforming to revolutionary paradigm.
• This technique enables precise genomic modifications in many different organisms and
tissues. Cas9 protein is an RNA guided endonuclease utilized for creating targeted
double-stranded breaks with only a short RNA sequence to confer recognition of the
target in animals and plants.
CRISPER CAS9:

• Development of genetically edited (GE) crops similar to those developed by conventional

or mutation breeding using this potential technique makes it a promising and extremely
versatile tool for providing sustainable productive agriculture for better feeding of rapidly
growing population in a changing climate.
 CAS9/SGRNA SYSTEM

• The basic strategy of Cas9/sgRNA system. The Cas9 is a

RNA guided endonuclease consists of two nuclease
domains namely HNH and RuvC. The target specificity of
Cas9 depends upon the guide sequence (20 nt) short
guide RNA (sgRNA). The target sites must lay
immediately 5`of a PAM (Protospacer Adjacent Motif)
sequence of the form N20-NGG (or N20-NAG). The
Cas9 nuclease induces double stranded breaks (DSB) at
the target site which can be repaired either by Non-
Homologous End Joining method or Homologous
Recombination by cellular system which results in gene
disruption by indels or gene addition/correction,
respectively
CURRENT FRAMEWORKS FOR GMO’S
IMPLEMENTATION:

• Process-oriented (triggered by the process by which

food and feed is derived)
• Product-oriented (triggered by the safety of product)
EU REGULATIONS AND FRAMEWORKS :

• The biggest fallacy with EU regulations is that they begin with the assumption that GM crops
are intrinsically different (i.e. more dangerous) from their non-GM equivalent.
• When GM regulations were formulated in the late 1980s, GM crops were not well known and
plant genomics was in its infancy.
• From the beginning the USA and Canada avoided this error.
• Today however, we know, even in Europe, that GM crops are not very different from the
parent crop, and with the emergence of NBTs, it has become clear that there is a continuous
spectrum of minor differences between them, such that it is impossible to draw a
differentiating line.. However, the Canadian (and partly the US) system is different in that it
considers the food safety of all new plant varieties, GM or traditional.
FRAMEWORKS USED IN THE USA AND CANADA:

• A modified organism is subject to regulatory oversight and risk assessment by the

respective authorities if the recipient organism or the source organism of the introduced
genetic elements or traits is known to display risky and/or unwanted characteristics (e.g.,
plant pathogenicity, weediness, toxicity or pesticidal effects or compositional differences
that are not generally recognized as safe).
• The product-oriented regulatory trigger used in Canada can also be considered risk-
oriented, which is triggered by the safety of product. Such a system makes more scientific
sense for improved food safety and consumer confidence.
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• Any
• Questions ?