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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.
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 large cell size), and the device was used to deliver particles coated with a marker gene which would relay a signal if proper insertion of the DNA transcript occurred. Genetic transformation was demonstrated upon observed expression of the marker gene within onion cells.
The earliest custom manufactured gene guns (fabricated by Nelson Allen) used a 22 caliber nail gun cartridge to propel a polyethylene cylinder (bullet) down a 22 caliber Douglas barrel. A droplet of the tungsten powder coated with genetic material was placed onto the bullet and shot down into a Petri dish below. The bullet welded to the disk below the Petri plate, and the genetic material blasted into the sample with a doughnut effect involving devastation in the middle of the sample with a ring of good transformation around the periphery. The gun was connected to a vacuum pump and was placed under a vacuum while firing. The early design was put into limited production by a Rumsey-Loomis (a local machine shop then at Mecklenburg Road in Ithaca, NY, USA).
Biolistics, Inc sold Dupont the rights to manufacture and distribute an updated device with improvements including the use of helium as a non-explosive propellant and a multi-disk collision delivery mechanism to minimize damage to sample tissues. Other heavy metals such as gold and silver are also used to deliver genetic material with gold being favored due to lower cytotoxicity in comparison to tungsten projectile carriers.
Biolistic transformation involves the integration of a functional fragment of DNA—known as a DNA construct—into target cells. A gene construct is a DNA cassette containing all required regulatory elements for proper expression within the target organism.[page needed] While gene constructs may vary in their design depending on the desired outcome of the transformation procedure, all constructs typically contain a combination a promoter sequence, a terminator sequence, the gene of interest, and a reporter gene.
Gene guns are mostly used with plant cells. However, there is much potential use in humans and other animals as well.
The target of a gene gun is often a callus of undifferentiated plant cells or a group of immature embryos growing on gel medium in a Petri dish. After the DNA-coated gold particles have been delivered to the cells, the DNA is used as a template for transcription (transient expression) and sometimes it integrates into a plant chromosome ('stable' transformation)
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Gene gun AI simulator
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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.
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 large cell size), and the device was used to deliver particles coated with a marker gene which would relay a signal if proper insertion of the DNA transcript occurred. Genetic transformation was demonstrated upon observed expression of the marker gene within onion cells.
The earliest custom manufactured gene guns (fabricated by Nelson Allen) used a 22 caliber nail gun cartridge to propel a polyethylene cylinder (bullet) down a 22 caliber Douglas barrel. A droplet of the tungsten powder coated with genetic material was placed onto the bullet and shot down into a Petri dish below. The bullet welded to the disk below the Petri plate, and the genetic material blasted into the sample with a doughnut effect involving devastation in the middle of the sample with a ring of good transformation around the periphery. The gun was connected to a vacuum pump and was placed under a vacuum while firing. The early design was put into limited production by a Rumsey-Loomis (a local machine shop then at Mecklenburg Road in Ithaca, NY, USA).
Biolistics, Inc sold Dupont the rights to manufacture and distribute an updated device with improvements including the use of helium as a non-explosive propellant and a multi-disk collision delivery mechanism to minimize damage to sample tissues. Other heavy metals such as gold and silver are also used to deliver genetic material with gold being favored due to lower cytotoxicity in comparison to tungsten projectile carriers.
Biolistic transformation involves the integration of a functional fragment of DNA—known as a DNA construct—into target cells. A gene construct is a DNA cassette containing all required regulatory elements for proper expression within the target organism.[page needed] While gene constructs may vary in their design depending on the desired outcome of the transformation procedure, all constructs typically contain a combination a promoter sequence, a terminator sequence, the gene of interest, and a reporter gene.
Gene guns are mostly used with plant cells. However, there is much potential use in humans and other animals as well.
The target of a gene gun is often a callus of undifferentiated plant cells or a group of immature embryos growing on gel medium in a Petri dish. After the DNA-coated gold particles have been delivered to the cells, the DNA is used as a template for transcription (transient expression) and sometimes it integrates into a plant chromosome ('stable' transformation)