Transfection By Agrobacterium
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Transfection By Agrobacterium
Transfection is the process of deliberately introducing naked or purified nucleic acids into Eukaryote, eukaryotic Cell (biology), cells. It may also refer to other methods and cell types, although other terms are often preferred: "transformation (genetics), transformation" is typically used to describe non-viral DNA transfer in bacteria and non-animal eukaryote, eukaryotic cells, including plant cells. In animal cells, transfection is the preferred term, as the term "transformation" is also used to refer to a cell's progression to a cancerous state (carcinogenesis). Transduction (genetics), Transduction is often used to describe virus-mediated gene transfer into prokaryotic cells. The word ''transfection'' is a portmanteau of the prefix ''trans-'' and the word "infection." Genetics, Genetic material (such as DNA supercoil, supercoiled plasmid DNA or siRNA constructs), may be transfected. Transfection of animal cells typically involves opening transient pores or "holes" in the pla ...
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Nucleic Acids
Nucleic acids are large biomolecules that are crucial in all cells and viruses. They are composed of nucleotides, which are the monomer components: a 5-carbon sugar, a phosphate group and a nitrogenous base. The two main classes of nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). If the sugar is ribose, the polymer is RNA; if the sugar is deoxyribose, a variant of ribose, the polymer is DNA. Nucleic acids are chemical compounds that are found in nature. They carry information in cells and make up genetic material. These acids are very common in all living things, where they create, encode, and store information in every living cell of every life-form on Earth. In turn, they send and express that information inside and outside the cell nucleus. From the inner workings of the cell to the young of a living thing, they contain and provide information via the nucleic acid sequence. This gives the RNA and DNA their unmistakable 'ladder-step' order of ...
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Cationic
An ion () is an atom or molecule with a net electrical charge. The charge of an electron is considered to be negative by convention and this charge is equal and opposite to the charge of a proton, which is considered to be positive by convention. The net charge of an ion is not zero because its total number of electrons is unequal to its total number of protons. A cation is a positively charged ion with fewer electrons than protons (e.g. K+ (potassium ion)) while an anion is a negatively charged ion with more electrons than protons (e.g. Cl− (chloride ion) and OH− (hydroxide ion)). Opposite electric charges are pulled towards one another by electrostatic force, so cations and anions attract each other and readily form ionic compounds. Ions consisting of only a single atom are termed ''monatomic ions'', ''atomic ions'' or ''simple ions'', while ions consisting of two or more atoms are termed polyatomic ions or ''molecular ions''. If only a + or − is present, it indicate ...
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Retrovirus
A retrovirus is a type of virus that inserts a DNA copy of its RNA genome into the DNA of a host cell that it invades, thus changing the genome of that cell. After invading a host cell's cytoplasm, the virus uses its own reverse transcriptase enzyme to produce DNA from its RNA genome, the reverse of the usual pattern, thus ''retro'' (backward). The new DNA is then retroviral integration, incorporated into the host cell genome by an integrase enzyme, at which point the retroviral DNA is referred to as a provirus. The host cell then treats the viral DNA as part of its own genome, transcribing and translating the viral genes along with the cell's own genes, producing the proteins required to assemble new copies of the virus. Many retroviruses cause serious diseases in humans, other mammals, and birds. Retroviruses have many subfamilies in three basic groups. * Oncovirus, Oncoretroviruses (cancer-causing retroviruses) include human T-lymphotropic virus (HTLV) causing a type of leuk ...
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Virus
A virus is a submicroscopic infectious agent that replicates only inside the living Cell (biology), cells of an organism. Viruses infect all life forms, from animals and plants to microorganisms, including bacteria and archaea. Viruses are found in almost every ecosystem on Earth and are the most numerous type of biological entity. Since Dmitri Ivanovsky's 1892 article describing a non-bacterial pathogen infecting tobacco plants and the discovery of the tobacco mosaic virus by Martinus Beijerinck in 1898, more than 16,000 of the millions of List of virus species, virus species have been described in detail. The study of viruses is known as virology, a subspeciality of microbiology. When infected, a host cell is often forced to rapidly produce thousands of copies of the original virus. When not inside an infected cell or in the process of infecting a cell, viruses exist in the form of independent viral particles, or ''virions'', consisting of (i) genetic material, i.e., long ...
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Lipofection
Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. It may also refer to other methods and cell types, although other terms are often preferred: "transformation" is typically used to describe non-viral DNA transfer in bacteria and non-animal eukaryotic cells, including plant cells. In animal cells, transfection is the preferred term, as the term "transformation" is also used to refer to a cell's progression to a cancerous state (carcinogenesis). Transduction is often used to describe virus-mediated gene transfer into prokaryotic cells. The word ''transfection'' is a portmanteau of the prefix ''trans-'' and the word "infection." Genetic material (such as supercoiled plasmid DNA or siRNA constructs), may be transfected. Transfection of animal cells typically involves opening transient pores or "holes" in the cell membrane to allow the uptake of material. Transfection can be carried out using calcium phosphate (i.e. trica ...
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Hydrostatic Pressure
Hydrostatics is the branch of fluid mechanics that studies fluids at hydrostatic equilibrium and "the pressure in a fluid or exerted by a fluid on an immersed body". The word "hydrostatics" is sometimes used to refer specifically to water and other liquids, but more often it includes both gases and liquids, whether Compressible flow, compressible or Incompressible flow, incompressible. It encompasses the study of the conditions under which fluids are at rest in Mechanical equilibrium, stable equilibrium. It is opposed to ''fluid dynamics'', the study of fluids in motion. Hydrostatics is fundamental to ''hydraulics'', the engineering of equipment for storing, transporting and using fluids. It is also relevant to geophysics and astrophysics (for example, in understanding plate tectonics and the anomalies of the Gravity of Earth, Earth's gravitational field), to meteorology, to medicine (in the context of blood pressure), and many other fields. Hydrostatics offers physical expl ...
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Impalefection
Impalefection is a method of gene delivery using nanomaterials, such as carbon nanofibers, carbon nanotubes, nanowires. Needle-like nanostructures are synthesized perpendicular to the surface of a substrate. Plasmid DNA containing the gene, and intended for intracellular delivery, is attached to the nanostructure surface. A chip with arrays of these needles is then pressed against cells or tissue. Cells that are impaled by nanostructures can express the delivered gene(s). As one of the types of transfection, the term is derived from two words – impalement and infection. Applications One of the features of impalefection is spatially resolved gene delivery that holds potential for such tissue engineering approaches in wound healing as gene activated matrix technology. Though impalefection is an efficient approach ''in vitro'', it has not yet been effectively used ''in vivo'' on live organisms and tissues. Carrier materials Vertically aligned carbon nanofiber arrays prepared by ...
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Gene Gun
In genetic engineering, a gene gun or biolistic particle delivery system is a device used to deliver exogenous DNA (transgenes), RNA, or protein to cells. By coating particles of a heavy metal with a gene of interest and firing these micro-projectiles into cells using mechanical force, an integration of desired genetic information can be introduced into desired cells. The technique involved with such micro-projectile delivery of DNA is often referred to as biolistics, short for "biological ballistics". This device is able to transform almost any type of cell and is not limited to the transformation of the nucleus; it can also transform organelles, including plastids and mitochondria. Gene gun design The gene gun was originally a Crosman air pistol modified to fire dense tungsten particles. It was invented by John C Sanford, Ed Wolf, and Nelson Allen at Cornell University along with Ted Klein of DuPont between 1983 and 1986. The original target was onions (chosen for their ...
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Microinjection
Microinjection is the use of a glass micropipette to inject a liquid substance at a microscopic or borderline macroscopic level. The target is often a living cell but may also include intercellular space. Microinjection is a simple mechanical process usually involving an inverted microscope with a Optical power, magnification power of around 200x (though sometimes it is performed using a dissecting stereo microscope at 40–50x or a traditional Optical microscope#Compound microscope, compound upright microscope at similar power to an inverted model). For processes such as cellular or pronucleus, pronuclear injection the target cell is positioned under the microscope and two micromanipulation, micromanipulators—one holding the pipette and one holding a microcapillary needle usually between 0.5 and 5 μm in diameter (larger if injecting stem cells into an embryo)—are used to penetrate the cell membrane and/or the nuclear envelope. In this way the process can be used to int ...
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Nanoparticle
A nanoparticle or ultrafine particle is a particle of matter 1 to 100 nanometres (nm) in diameter. The term is sometimes used for larger particles, up to 500 nm, or fibers and tubes that are less than 100 nm in only two directions. At the lowest range, metal particles smaller than 1 nm are usually called atom clusters instead. Nanoparticles are distinguished from microparticles (1-1000 μm), "fine particles" (sized between 100 and 2500 nm), and "coarse particles" (ranging from 2500 to 10,000 nm), because their smaller size drives very different physical or chemical properties, like colloidal properties and ultrafast optical effects or electric properties. Being more subject to the Brownian motion, they usually do not sediment, like colloid, colloidal particles that conversely are usually understood to range from 1 to 1000 nm. Being much smaller than the wavelengths of visible light (400-700 nm), nanoparticles cannot be seen with ordinary ...
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Cell (biology)
The cell is the basic structural and functional unit of all life, forms of life. Every cell consists of cytoplasm enclosed within a Cell membrane, membrane; many cells contain organelles, each with a specific function. The term comes from the Latin word meaning 'small room'. Most cells are only visible under a light microscope, microscope. Cells Abiogenesis, emerged on Earth about 4 billion years ago. All cells are capable of Self-replication, replication, protein synthesis, and cell motility, motility. Cells are broadly categorized into two types: eukaryotic cells, which possess a Cell nucleus, nucleus, and prokaryotic, prokaryotic cells, which lack a nucleus but have a nucleoid region. Prokaryotes are single-celled organisms such as bacteria, whereas eukaryotes can be either single-celled, such as amoebae, or multicellular organism, multicellular, such as some algae, plants, animals, and fungi. Eukaryotic cells contain organelles including Mitochondrion, mitochondria, which ...
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Bacteriophage
A bacteriophage (), also known informally as a phage (), is a virus that infects and replicates within bacteria. The term is derived . Bacteriophages are composed of proteins that Capsid, encapsulate a DNA or RNA genome, and may have structures that are either simple or elaborate. Their genomes may encode as few as four genes (e.g. Bacteriophage MS2, MS2) and as many as hundreds of genes. Phages replicate within the bacterium following the injection of their genome into its cytoplasm. Bacteriophages are among the most common and diverse entities in the biosphere. Bacteriophages are ubiquitous viruses, found wherever bacteria exist. It is estimated there are more than 1031 bacteriophages on the planet, more than every other organism on Earth, including bacteria, combined. Viruses are the most abundant biological entity in the water column of the world's oceans, and the second largest component of biomass after prokaryotes, where up to 9x108 virus, virions per millilitre have b ...
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