Transgenic animals are genetically engineered to contain genes from another species. The first transgenic animal was produced by microinjecting DNA into fertilized mouse eggs. This allows the new genes to integrate into the genome and be passed to offspring. Knockout mice have a specific endogenous gene altered so it is no longer expressed normally. They are used to study gene function and model human diseases. Dolly the sheep was the first mammal cloned from an adult cell, showing that nuclear transfer can generate a live offspring genetically identical to the donor animal.
it contain some production techniques of transgenic animals with some examples and utility in drug development (available transgenic animals model of drug and their activity).
Applications and uses in different field
Another techniques like transposons and knock-out & knock-in discussed later
Transgenic animal production and its applicationkishoreGupta17
A genetically modified animal with the heterologous gene of interest being inserted for the purpose of biopharming or make a diseased model to study the consequences of disease and its probable therapy
Refers to an animal in which there has been a deliberate modification of the genome - the material responsible for inherited characteristics - in contrast to spontaneous mutation.
Foreign DNA is introduced into the animal, using recombinant DNA technology,
it contain some production techniques of transgenic animals with some examples and utility in drug development (available transgenic animals model of drug and their activity).
Applications and uses in different field
Another techniques like transposons and knock-out & knock-in discussed later
Transgenic animal production and its applicationkishoreGupta17
A genetically modified animal with the heterologous gene of interest being inserted for the purpose of biopharming or make a diseased model to study the consequences of disease and its probable therapy
Refers to an animal in which there has been a deliberate modification of the genome - the material responsible for inherited characteristics - in contrast to spontaneous mutation.
Foreign DNA is introduced into the animal, using recombinant DNA technology,
Introduction
History
Landmarks Events in Transgenic Livestock Research
Techniques/ Method for Gene Transfer
Examples of transgenesis
Importance
Application
Limitation
Issue related to Transgenic Technology
Ethical concerns and how to Overcome
Introduction.
Definition.
Importance of transgenic animals.
Transgenic mice
Methods for introducing a foreign gene:
The retroviral vector method
The DNA microinjection method/ pronuclear microinjection
Genetically engineered embryonic stem cells
Transgenic fish
What is transgenic fish?
A few facts to know to know about transgenic fish.
Important points needed for genetic engineering (gene transfer) to produce transgenic fish.
Development of transgenic fishes.
A few examples
Auto-transgenesis.
Controlled culture of transgenic fish and feed.
Gene transfer technology for development of transgenic fishes.
Gene flow.
Food safety issues.
Conclusion.
Bibliography.
This presentation gives a comprehensive detail of transgenic animal, processes involve in the production of transgenic animal and also highlights several benefits of transgenic animal
A knockout mouse is a mouse in which a specific gene has been inactivated or“knocked out” by replacing it or disrupting it with an artificial piece of DNA.
The loss of gene activity often causes changes in a mouse's phenotype and thus provides valuable information on the function of the gene.
Introduction
History
Landmarks Events in Transgenic Livestock Research
Techniques/ Method for Gene Transfer
Examples of transgenesis
Importance
Application
Limitation
Issue related to Transgenic Technology
Ethical concerns and how to Overcome
Introduction.
Definition.
Importance of transgenic animals.
Transgenic mice
Methods for introducing a foreign gene:
The retroviral vector method
The DNA microinjection method/ pronuclear microinjection
Genetically engineered embryonic stem cells
Transgenic fish
What is transgenic fish?
A few facts to know to know about transgenic fish.
Important points needed for genetic engineering (gene transfer) to produce transgenic fish.
Development of transgenic fishes.
A few examples
Auto-transgenesis.
Controlled culture of transgenic fish and feed.
Gene transfer technology for development of transgenic fishes.
Gene flow.
Food safety issues.
Conclusion.
Bibliography.
This presentation gives a comprehensive detail of transgenic animal, processes involve in the production of transgenic animal and also highlights several benefits of transgenic animal
A knockout mouse is a mouse in which a specific gene has been inactivated or“knocked out” by replacing it or disrupting it with an artificial piece of DNA.
The loss of gene activity often causes changes in a mouse's phenotype and thus provides valuable information on the function of the gene.
Powerpoint accompanying research paper on the Teratogenic effects of Fipronil (GABAergic antagonist) on the neurological development of zebrafish (as model of vertebrates).
stem cell research is yet to be advanced , once fully developed can alleviate human suffering, this ppt reviews the contemporary evidence, pitfalls and challenges
description of transgenic animals and production with desired traits using different methods and their applications and their advantages and disadvantages
This presentation aims to provide an in-depth understanding of the science behind creating transgenic animals, explore their potential applications, and delve into the ethical considerations surrounding this emerging field of research.
Definition and Background:
We begin by defining transgenic animals as organisms that have had their genetic material intentionally altered through the introduction of foreign genes. This groundbreaking field of genetic engineering has its roots in the development of recombinant DNA technology in the 1970s, which enabled the transfer of genes across different species.
Genetic Engineering Techniques:
This section delves into the techniques employed to create transgenic animals, emphasizing the following key methodologies:
a. DNA Microinjection: The introduction of foreign DNA into the pronucleus of a fertilized embryo, allowing the foreign gene to be incorporated into the animal's genome and expressed in its cells.
b. Gene Targeting: The precise modification of an organism's genome by replacing or disrupting specific genes using technologies such as homologous recombination or CRISPR-Cas9.
c. Somatic Cell Nuclear Transfer (SCNT): The cloning technique involving the transfer of a nucleus from a somatic cell into an enucleated egg, resulting in the creation of an embryo with the same genetic makeup as the somatic cell donor.
Applications of Transgenic Animals:
This section explores the wide-ranging applications of transgenic animals across various fields, including:
a. Biomedical Research: Transgenic animals serve as invaluable models for studying human diseases and testing potential therapies, enabling significant advancements in medical research.
b. Agriculture: Transgenic animals can be engineered to possess desirable traits, such as increased resistance to diseases or improved meat quality, offering the potential to enhance agricultural productivity and sustainability.
c. Pharmaceutical Production: Transgenic animals can be designed to produce therapeutic proteins or antibodies in their milk or blood, providing a cost-effective means of manufacturing valuable pharmaceutical products.
d. Organ Transplantation: Research on transgenic animals has explored the possibility of generating organs that are genetically compatible with humans, addressing the shortage of donor organs for transplantation.
Transgenesis is the future of healthcare where the world is focusing on it so why not us? Let's delve into the exclusive depth of this transgenesis in the slide.
the following file contains information regarding the research based on transgenic animals. It is a biotechnological approach and an assignment(report) of a student of B.S.C second-year biotechnology.
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.
Toxic effects of heavy metals : Lead and Arsenicsanjana502982
Heavy metals are naturally occuring metallic chemical elements that have relatively high density, and are toxic at even low concentrations. All toxic metals are termed as heavy metals irrespective of their atomic mass and density, eg. arsenic, lead, mercury, cadmium, thallium, chromium, etc.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
The ability to recreate computational results with minimal effort and actionable metrics provides a solid foundation for scientific research and software development. When people can replicate an analysis at the touch of a button using open-source software, open data, and methods to assess and compare proposals, it significantly eases verification of results, engagement with a diverse range of contributors, and progress. However, we have yet to fully achieve this; there are still many sociotechnical frictions.
Inspired by David Donoho's vision, this talk aims to revisit the three crucial pillars of frictionless reproducibility (data sharing, code sharing, and competitive challenges) with the perspective of deep software variability.
Our observation is that multiple layers — hardware, operating systems, third-party libraries, software versions, input data, compile-time options, and parameters — are subject to variability that exacerbates frictions but is also essential for achieving robust, generalizable results and fostering innovation. I will first review the literature, providing evidence of how the complex variability interactions across these layers affect qualitative and quantitative software properties, thereby complicating the reproduction and replication of scientific studies in various fields.
I will then present some software engineering and AI techniques that can support the strategic exploration of variability spaces. These include the use of abstractions and models (e.g., feature models), sampling strategies (e.g., uniform, random), cost-effective measurements (e.g., incremental build of software configurations), and dimensionality reduction methods (e.g., transfer learning, feature selection, software debloating).
I will finally argue that deep variability is both the problem and solution of frictionless reproducibility, calling the software science community to develop new methods and tools to manage variability and foster reproducibility in software systems.
Exposé invité Journées Nationales du GDR GPL 2024
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show that a plume deposit from a powerful eruption at Pillan Patera has covered part
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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.
2. Transgenic Animals: A Focus
on Transgenic Mice Studies
http://www.hku.hk/biochem/tgcentre/transcentre.html
3. Introduction
Transgenic animals:
– Animals which have been genetically engineered to
contain one or more genes from an exogenous source.
– Transgenes are integrated into the genome.
– Transgenes can be transmitted through the germline to
progeny.
– First transgenic animal produced = “Founder Animal”
4. Introduction of foreign genes
into intact organisms
Procedure is basically the same regardless of
which animal is involved.
Integration usually occurs prior to DNA
replication in the fertilized oocyte.
– Majority of transgenic animals carry the gene in all of
their cells, including the germ cells. Transmission to
next generation requires germline integration.
– Some integration events occur subsequent to DNA
replication giving rise to mosaic animals which may
or may not contain the transgene in its germline.
6. First Breeding Pair:
– Fertile male + superovulated female
• Fertile male = stud (changed regularly to ensure
performance)
• Superovulated female = immature female induced to
superovulate
– Pregnant mare’s serum (=FSH) on day 1
– Human Chorionic Gonadotropin (=LH) on day 3
• Mated on day 3
• Fertilized oocytes microinjected on day 4 with foreign DNA
construct.
• Microinjected oocytes are transferred to the oviducts of
surrogate mothers at end of day 4.
7. Second breeding pair:
– Sterile male + surrogate mother
• Sterile male produced through vasectomy
• Surrogate mother must mate to be suitable recipient of
injected eggs
• Mated on day 3
• Microinjected oocytes from first breeding pair are
transferred to oviducts on day 4
• Embryos implant in uterine wall and are born 19 days later.
• Southern blotting techniques confirm presence and copy
number of transgenes.
9. Third breeding pair:
– Foster parents
• Fertile male + female mated to give birth on same
day surrogate mother
• Serves as foster parent if caesarian section is
required for surrogate mother
15. Integration of Transgene into
One Chromosome
Normally the transgene inserts into one
chromosome giving rise to a heterozygote.
– 50% probability of passing transgene onto offspring.
Two heterozygous mice may be bred to obtain a
homozygous line that contains the transgene on
both chromosomes.
– 100% probability of passing transgene onto offspring.
Most transgenes are stably transmitted for many
generations without detectable rearrangement.
16. Mechanisms of DNA
Integration
Linear molecules integrate more efficiently than circular
molecules (~5x)
Once in the oocyte, the linear molecules circularize.
Usually all of the molecules that integrate are on the same
chromosome and at the same site.
Multiple copies are usually arranged in a tandem, head-
to-tail array.
The size of the DNA molecule (0.7 – 50Kb) is not an
important parameter.
The concentration and purity of the injected DNA is
critical (1-3 µg/ml maximum).
17. Working Hypothesis of DNA
Integration
The ends of the injected linear DNA integrate at breaks that occur
spontaneously in the chromosome.
Other injected molecules which have circularized probably
recombine with each other and the integrated copies to generate a
tandem, head-to-tail array.
Recombination is probably favored because of high local DNA
concentration and special properties such as the absence of normal
chromatin structure.
The number of chromosomal breaks is presumably limiting
explaining the low number of integration events and why different
DNA molecules are usually integrated at the same site.
18. Gene Expression in
Transgenic Mice
In order to discriminate the products of the
injected gene from those of an endogenous
counterpart, the injected gene must be marked in
some way.
– Mini-genes where exons are deleted of cDNA where
introns are absent.
– Modification by insertion/deletion/mutagenesis of a
few nucleotides (e.g. the gain or loss of a restriction
endonuclease site).
– Hybrid genes where foreign epitopes are expressed on
transgenic products.
19. Tissue-Specific Gene
Expression
Generally, if a tissue-specific gene is expressed at all,
then it is expressed appropriately, despite the fact that it
has integrated at a different chromosomal location.
– Trans-acting proteins involved in establishing tissue-specific
expression are capable of finding their cognate sequences and
activation transcription at various chromosomal locations.
– Levels of expression vary between founder animals as
chromosomal position can influence accessibility of the
transgenes to these trans-acting transcription factors.
– Some founders do not express the transgene at all owing to
integration into heterochromatin domains where DNA is
methylated heavily (silent).
20. Prokaryotic Sequences Must be
Removed for Optimal Expression
Prokaryotic sequences derived from the plasmid or
bacteriophage vector used for replication of the transgene
in bacteria can be inhibitory or “poisonous” for some
transgenes.
Therefore, the transgene fragment requires purification
from contaminating vector sequence prior to
microinjection.
21. Possible Reasons for Lack of
Transgene Expression
Integration in cis-acting silencer sequences (the negative counterpart
of enhancer elements) might be sites for covalent modification of
DNA (e.g. methylation) which might initiate condensation into an
inactive chromatin configuration, or they might phase nucleosomes
in an inappropriate manner.
The inadvertent loss of certain regulatory sequences during the
production of the constructs (e.g. topoisomerase-binding sites,
nuclear matrix-attachment sites).
Use of cDNA rather than genomic DNA. (Introns thought to
contribute to stability of mRNA and may even contain enhancer
sequences essential for tissue-specific expression. Flanking DNA
may also contain regulatory sequences.)
22. Examples of Studies Utilizing
Transgenic Mice
The Oncomouse
– c-myc oncogene + MMTV sequences breast cancer
– Int-2 oncogene + viral promoter prostate cancer
To obtain abnormal expression of genes to study their
effects
– Rat growth hormone + cadmium-inducible metallothionein
promoter
– Transgenic mouse was much larger, but also suffered
complications with fertility and organ diseases
23. To study developmentally regulated genes
http://www.ucihs.uci.edu/anatomy/calofpix1b.html
24. More Examples of Studies
Utilizing Transgenic Mice
“Pharm” animals (transgenic livestock)
– Bioreactors whose cells have been engineered to
synthesize marketable proteins
– DNA constructs contain desired gene and appropriate
regulatory sequences (tissue-specific promoters)
– More economical than producing desired proteins in
cell culture
25. Examples of Bioreactors
Naked human Hb from
pigs
Human lactoferrin in
cows’ milk
Alpha-1-antitrypsin in
sheep
HGH in mouse urine
(uroplakin promoters)
Human antibodies in mice
(H and L chain tgenics
hybridomas)
CfTCR in goats
Tissue plasminogen
activator (TPA) in goats
Human antithrombin III
in goats
Malaria antigens in goats
(vaccine)
Alpha-glucosidase in
rabbits (Pompe’s disease
29. Transgenic Pigs for the Production
of Organs for Transplantation
Pig organs are rejected acutely due to the
presence of human antibodies to pig antigens.
Once human antibodies are bound to pig organs,
human complement is activated and triggers the
complement cascade and organ destruction.
Transgenic pigs with complement inhibitors have
been produced and are gaining feasibility as a
source of xenogeneic organs for transplantation.
34. What is a Knockout Mouse?
A really good-looking mouse?
A mouse in which a very specific
endogenous gene has been altered in such
a way that interferes with normal
expression, i.e. it has been knocked out.
35. Why Produce KO Mice?
To study effects of gene products,
biochemical pathways, alternative
(compensatory) pathways, and
developmental pathways
To recreate human diseases in animals to
establish models to test the beneficial
effects of drugs or gene therapy.
36. Procedure for Generating
a KO Mouse
Gene alteration in KO mice is targeted to very specific
genes.
DNA must integrate at precise positions in the genome.
Integration of the altered gene takes place in embryonic
stem cells ex vivo.
Verification of exact location of integration occurs before
the ESC is introduced into blastocysts to become part of
the developing embryo.
37. Pluripotent ES Cells
Pluripotent ES cells are undifferentiated early embryonic
cells derived from the inner cell mass of mouse
blastocysts.
In vitro ES cells must be grown on a feeder layer of
fibroblasts to prevent them from differentiating.
Introduction of the transgene is achieved by
electroporation of retroviral infection.
The transgene must integrate via recombination, not
randomly.
Cells transfected successfully can be identified prior to
injection into blastocysts.
38. Specific Gene Targeting in ES
Cells
Gene targeting can be achieved using gene
constructs designed for homologous
recombination. This technique can be used to
either:
– Knockout functional genes to study their contribution
to different developmental or disease processes (null
mutations)
• Genes encoding β2m, MHC class I and II. CD2, Ii, TCR, Ig,
IL-4, IL-2, FcεR, TAP1/2, RAG-2,and many more (>100)!
– Replace a functional gene for a mutated/non-
functional gene to restore wild type phenotype .
• Gene encoding HGPRT in mice deficient for HGPRT (called
Lesch-Nhyan syndrome in humans).
39. DNA Constructs for
Recombination
DNA vectors contain the gene of interest which
has been interrupted with an antibiotic resistance
gene (hygromycin resistance, or G418
resistance).
To ensure targeted integration has occurred, the
flanking DNA contains the thymidine kinase
gene. If TK integrates (random insertion), then
the transfected cells die when grown in selective
media (gancyclovir).
40.
41. Selection of Targeted ES Cells
Gancyclovir resistant and G418 resistant ES cells
grow into small clumps on top of feeder cells.
The colonies of cells can be “picked” off and
transferred to new wells (at 0.3 cells per well
seeding density) containing feeder cells.
When sufficient numbers of cells are obtained,
they are:
– Frozen for safe storage
– Analyzed by Southern blotting or PCR to determine
nature of integration event
– Microinjected into the blastocoel cavity of blastocysts.