Transgenic animals

Transgenic animals can be described as animals that have altered or modified genomes. In
transgenic animals, a foreign gene is injected into the genome intending to improve the genetic
traits of the targeted animal. In the beginning, genetic traits were only improved by using selective
breeding methods. In this technique, selective breeding methods were taken into consideration to
improve the traits of the targeted animal. However, this technique was costly and time-consuming;
hence it was substituted by recombinant DNA technology.

Transgenesis is a method where a foreign gene with all the diverse traits is initiated into the target
animal’s genome. This newly introduced foreign gene is termed the transgene, whereas a
transgenic is the animal whose genome is altered. Once it’s in the targeted animal, it is passed to
the coming generations. Apart from being transgenic, these animals are also genetically modified
organisms.

Methods for Creating transgenic animals

There are several methods through which transgenic animals can be created. These include-
• Physical Transfection
In the physical transfection method, the interested gene is directly injected into the pronucleus of
the fertilized ovum. This method was widely used in mammals and was proven as the first method
is to be highly effective. Ultrasound, electroporation, and particle bombardment are other popular
physical transfection methods.
• Chemical Transfection
Gene transfection can also be performed through one of the chemical methods. While performing
this method, in the presence of calcium phosphate, the targeted DNA is taken as a co-precipitate
that allows the DNA uptake. The cells of mammals can take foreign DNA from the medium of
culture.
• Retrovirus-Mediated Gene Transfer
The gene gets transferred through a vector to multiply the expression changes since the
retroviruses can infect the entire host cell. To inject the interested genes into the targeted animal,
they are used as vectors.
• Viral Vectors
Viruses play a significant role in transferring DNA into animal cells. These viruses have the ability
to infect the host cell, replicate efficiently, and express well.
• Bactofection
Bactofection is when the desired gene is transferred to the target through bacteria.

Applications of transgenic animals

Let’s discuss the applications of the transgenic animals-
• Normal Physiology and Development
As you already know, transgenic animals consist of a foreign gene, because of which the growth
factors get altered. Hence, these highly affect the body’s overall functioning and facilitate the gene
regulation study.
 • Study of Diseases
The primary reason behind the development of transgenic animals is to study. Further, the role
played by the genes to develop certain diseases. These are of great help as they are used in the
research centres for developing medicines.
• Biological Products
Transgenic animals produce several biological products, including nutritional supplements and
medicines. Several diseases like hereditary emphysema and phenylketonuria are treated using
these products,
• Vaccine Safety
The transgenic animals act as the models to test several vaccines to check their reliability and
safety before they get injected into human beings.

Conclusion
Till now, we have understood what transgenic animals are and what they are used for. Transgenic
animals are modified through biotechnology by inserting the desired DNA into the animals to be
modified. Animals are modified to study about various diseases, normal physiology and
development, check vaccine safety and form certain biological products.
 Knockout mouse

A knockout mouse is a genetically modified mouse that has a gene inactivated or removed from its
genome. The goal of creating knockout mice is to deactivate a gene to study its effect on the body.
Knockout mice are important animal models for studying the role of genes that have been
sequenced but whose functions have not been determined. Researchers can infer the function of a
gene by observing the differences in the mouse's behavior or physiology.

Knockout mice have been used to study and model:

Cancer, Obesity, Heart disease, Diabetes, Arthritis, Substance abuse, Anxiety, Aging, and
Parkinson's disease.
The technique for creating knockout mice involves:
1. Isolating the target gene
2. Engineering a modified DNA sequence
3. Introducing this into embryonic stem cells
4. Implanting the modified stem cells into mouse blastocysts.
5. This generates chimeric mice that can pass the modified gene to offspring.

Knockout strategies can be further subcategorized as:

• Constitutive knockout: Inactivates the gene of interest in all cells and tissues permanently
throughout development
• Conditional knockout: Allows for tissue- and/or stage-specific inactivation.
 Transgenic plants

Transgenic plants definition includes those plants which have one or more genes from another
species introduced into their genome through genetic engineering processes. The biolistic method,
in which a heavy metal coated with plasmid DNA is shot into cells, and Agrobacterium
tumefaciens-mediated transformation is two techniques.

Transgenic plants include maize, rice, brinjal, cabbage, cauliflowers, potato, and tomato. The goal
is to give the plant a new characteristic that does not arise naturally in the species. In this article,
we will study more about the development of transgenic plants and various methods used for their
development.
Transgenic Plants
• Transgenic plants are those that are created by inserting foreign DNA into a cell and
regenerating a plant from the cell.
• Genetic engineering techniques are used to change the DNA of transgenic plants.
• The goal of introducing a gene combination into a plant is to increase its productivity and
usefulness.
• This technique offers benefits like extended shelf life, increased yield, improved quality,
pest resistance, heat, cold and drought tolerance, as well as resistance to a range of biotic
and abiotic challenges.
• Additionally, foreign proteins of commercial and medicinal value can be expressed in
transgenic plants.
• Golden rice is one of the transgenic plants. The rice plant, which makes beta carotene, is
given the beta-carotene gene. The most common source is carrots, which when added to
rice plants begin producing beta-carotene. Golden rice is fortified with vitamin A to
increase its nutritional value because beta carotene is one of the precursors of vitamin A.
• Bt-cotton is an additional form of the transgenic plant. The Bt-gene originates from
bacteria and can be inserted into the cotton plant. The Bt-toxin, which is produced by this
gene and is not hazardous to people, kills the bugs that feed on cotton plants.

Development of Transgenic Plants

In a lab, the genetic makeup of a plant is changed, typically by adding one or more genes from its
genome, to create genetically modified plants. The new transgenic DNA is intended for the plant
cell’s nucleus. The biolistic method (particle gun method) or Agrobacterium tumefaciens-mediated
transformation method is used to create the majority of genetically modified plants.

1. Biolistic Method
Gene gun delivery of macromolecules DNA, RNA and proteins to the target cell involves the
coating of macromolecules with micro-carrier particles such as gold and tungsten. They are then
introduced into the target cell with high velocity, using an electric pulse or pressurised helium
pulse. Such technique involved with the delivery of microprojectile is referred to as biolistic.

The major limitation of this technique is that the DNA is randomly introduced into the target cell.

2. Agrobacterium Mediated Transformation Method

The “agrobacterium” approach is the following technique for creating genetically modified plants.
Agrobacterium tumefaciens, a soil-dwelling bacterium, is used in this process. It has the capacity
to spread a piece of its DNA into plant cells. Through a tumour-inducing plasmid, the DNA
fragment that infects the plant gets incorporated into a plant chromosome (Ti plasmid). The plant’s
cellular machinery may be controlled by the Ti plasmid, which can then exploit it to produce
numerous copies of its own bacterial DNA. A sizable circular DNA particle called the Ti plasmid
replicates separately from the bacterial chromosome.

This plasmid is significant because it has areas of transfer DNA (t DNA) where a researcher can
introduce a gene that can be transferred to a plant cell using a method called “floral dip.” A floral
dip entails dipping flowering plants into an agrobacterium solution that contains the desired gene
and then harvesting the transgenic seeds straight from the plant.

This strategy is advantageous since it is regarded as a more acceptable technique due to its natural
manner of transfer. Additionally, “agrobacterium” is extremely effective at moving big DNA
pieces. The fact that not all significant food crops can be infected by these bacteria is one of the
agrobacterium’s main drawbacks. This approach works exceptionally well for the dicotyledonous
plants like potatoes, tomatoes and tobacco plants.

Importance of Transgenic Plants

Insecticidal Resistance: A number of insect pests are harmful to the bacterium Bacillus
Thuringiensis. It creates a protein toxin that is the mediator of its fatal impact. The toxin gene can
be delivered directly into the plant’s genome using recombinant DNA techniques where it is
produced and protects the plant from insect pests.

Virus Resistant Plants: By adding viral coat proteins, TMV-resistant tomato and tobacco plants
can be created. Other viral-resistant transgenic plants include:
• Potato virus-resistant plants
• Rice that is resistant to RSV
• Black gram that is resistant to YMV
• Green gram that is resistant to YMV

Herbicidal Resistance: Herbicide-resistant plants are those that can withstand herbicides. Many
broad-range herbicides contain glyphosate as an active component. To produce glyphosate-
resistant transgenic tomato, potato, tobacco, and cotton, the aroA gene from Salmonella
typhimurium is inserted into a glyphosate EPSP synthase. To create tobacco plants resistant to E.
coli sulphonylurea, the mutant ALS (acetolactate synthase) gene from Arabidopsis is transformed.
To create atrazine-resistant transgenic plants, QB protein of photosystem II from mutant
Amaranthus hybrids is introduced into tobacco and other crops.

Nutritional Benefits: Every year, 500,000 kids lose their vision completely or partially due to
vitamin A deficiency. For a sizable portion of the world’s population, milled rice serves as their
main source of nutrition.

Crops with a high concentration of vitamin A cannot be produced using traditional breeding
techniques. Three genes, two from daffodils and one from a microbe, have been inserted into rice
by researchers. Transgenic rice has yellow-coloured seeds and produces more beta-carotene, which
is a precursor to vitamin A. Such golden or yellow rice may be an effective remedy for the issue of
vitamin A insufficiency in young infants living in tropical regions.
Conclusion
Genetic Modification is a technology that involves inserting DNA into an organism's genome.
New DNA is transferred into plant cells to create a genetically modified plant. Typically, the cells
are grown in tissue culture and eventually develop into plants. The new DNA will be passed down
to the seeds produced by these plants. Transgenesis allows for the enhancement of nutrients in
animal products, including their quantity, overall quality, and specific nutritional composition.
Transgenic technology could be used to transfer or enhance nutritionally beneficial traits.

Bt crops
Bt crops are transgenic crops that are genetically engineered from the DNA of bacterium Bacillus
thuringiensis.” Bt Crops are transgenic crops that produce the same toxin as the bacterium Bacillus
thuringiensis in the plant cell, thereby, protecting the crops from pests.
or
Bt plants, or genetically modified crops, are plants that contain DNA from the Bacillus
thuringiensis (Bt) bacterium. These plants produce Bt proteins, or toxins, that only target specific
insects. The toxins produced by the plant are called "plant-incorporated protectants".
Bt plants are effective at combating pests such as: European corn borer, Rootworm, Corn earworm,
and Bollworm. When an insect feeds on the transgenic plants, the toxic cry protein present in the
plants crystallizes the digestive system of insects, eventually leading to its death. However, it has
no harmful effects on the human digestive system.
The Environmental Protection Agency (EPA) in the USA approved the commercial production and
distribution of Bt crops in 1995. The use of Bt corn and Bt cotton result in less risk to human
health and the environment than chemical alternatives. Some examples of Bt crops
include: Cotton, Brinjal, Corn, Cry1Ab and Cry1F Bt corn, and Cry1Ac Bt cotton.

About Bt
Bt is a bacterium Bacillus thuringiensis which is a family of proteins originating from strains of the
bacterium Bt. To affect the different types of insects there are more than 200 different types of Bt
toxins. Basically, Bt is a spore-forming bacterium that produces crystals protein known as cry
protein which is toxic to many species of insects. It is found almost everywhere in the world. It is
also found in all types of terrain, including beaches, deserts, and tundra habitats. It is more
significantly used in agriculture, especially in organic farming.

It is also used in organic spraying programs and in transgenic crops. Bt is used to cause mortality
in pests. The toxin of Bt gets dissolved in the high pH insect gut and becomes active. The
toxins which were active attack the gut cells of the insect, punching holes in the lining. The spores
of Bt dribbles out of the gut and germinate in the insect causing the death of the insect within a
couple of days.

Bt Gene
Bt gene is a gene that is present in the bacterium of Bacillus thuringiensis which is a soil-dwelling
bacterium. It naturally produces a toxin that is fatal to certain herbivorous insects. Since 1920, it
has been utilized in organic farming in the form of insecticide spray. It is also a source of the gene
used to genetically modify a number of crops so that they produce the toxin on their own to
destroy various insect pests. Bt genes were first used by the Belgian company Plant Genetic
Systems in the year 1985 to develop genetically modified crops with insect tolerance by expressing
cry genes from Bt.

Introduction to Bt Crops

The full form of bt is Bacillus thuringiensis. It is a bacteria that naturally produces a crystal protein
that is toxic to many pest insects. These are the crops that are genetically engineered to produce the
same toxin as Bt in every cell of the plant so that pests do not destroy the plant. Bt crops are often
found safe because Bt is a common organic form of insecticide. When it is used by farmers it
degrades within a week and sometimes just in one day. The widespread use of Bt crops, pests
became bt resistance. This has become a greater threat to organic agriculture.

Types of Bt Crops
The types of Bt crops are as follows:
• Bt Cotton - The cotton plant is genetically modified with the Bt gene to protect the plants
from bollworm which is a major pest of cotton. Bt cotton is a transgenic crop that is an
insect-resistant designed to combat the bollworm. It was created by genetically altering the
cotton genome to express a microbial protein from the bacterium Bacillus thuringiensis.
The genetically modified gene which has been inserted into the plant's genome produces
toxin crystals that the plant would not normally produce at all. When this is ingested by a
certain population of organisms it dissolves within the gut lining which leads to the
organism's death.
• The worms which are present on the leaves of Bt cotton become lethargic and sleepy
resulting in less damage to the plants. The toxins which are produced by the crops are
ingested by the pests which result in their death.
• Bt Brinjal - It is also generated by the genetic transformation of a crystal protein gene cry
1 Ac from the bacterium Bacillus thuringiensis. This crop was developed to provide
resistance against insects called Lepidopteron. The proteins which are produced by Bt
genes bind to the receptors present on the insect's membrane which forms pores on the
membranes. This damages the digestive process and leads to the death of the insect.
• Bt Maize - The Bt maize was introduced to kill corn rootworm, Diabrotica virgifera which
is also known as a billion-dollar bug. These crops are also derived from the Bacillus
thuringiensis so the common name is Bt maize. However, this crop has revolutionized pest
control in a number of countries.