How does luciferase transfection work?

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Luciferase transfection is a method in genetic engineering whereby luciferase, a complex nucleic acid involved in a type of light-production in microorganisms and animals called bioluminescence, is introduced into an organism that does not naturally produce bioluminesce. The results of luciferase transfection include a potentially wide range of species of plants and animals, such as trees and pets, that “glow” in the dark.

Bioluminescence is most commonly observed in fireflies, as well as in certain species of anglerfish. It results from the activation of luciferin, a biological pigment that emits light when it oxidizes in the presence of luciferase. As a protein enzyme, luciferin acts as a catalyst for the production of light-emitting oxyluciferin, which otherwise proceeds much too slowly to produce normal visible light. There are a number of different types of luciferin, existing as small-molecule substrates for their corresponding protein enzyme luciferase.

Different types of organisms rely on different types of luciferin-luciferase pairs to emit their glow. The roughly 2,000 species of fireflies, comprising the Lampyridae family of beetles, contain a type of luciferin that uses the coenzyme nucleotide adenosine triphosphate (ATP), the main chemical responsible for storing and transporting chemical energy in animal and plant cells. The freshwater snail Latia neritoides uses a luciferin chemically known as (E)-2-methyl-4-(2,6,6-trimethyl-1-cyclohex-1-yl)-1-buten-1-ol formate. Certain types of squid and fish , meanwhile, carry luciferin inside a certain bacteria found in specialized light-producing tissues. Coelenterazine is a luciferin found in wide range of aquatic animals, ranging from tiny radiolarians, to centenophores, cnidarians, squid, copepods, chaetognaths, and certain species of fish and shrimp. It usually produces light in the blue range, along with vargulin, found in certain crustaceans and a fish called Porycthis. The familiar dynoflagellates, familiar for producing much of the phenomenon of night time phosphorescence, rely on a chlorophyll-derived luciferin.

In the laboratory, transfection involves the isolation of luciferase DNA derived from a particular species and its insertion into another species. Researchers have devised a variety of procedures for transfecting different types of luciferase to create genetically modified luciferin-producing organisms. Regardless of the particular technique, the parent DNA takes root in the cells of the host organism, where it is replicated and passed to daughter cells. Thus the genetic constitution of the cell line or organism is changed, and the organism can produce offspring with the modified DNA.

Mice, silkworms and potatoes are among the first of many organisms that have been engineered through transfection to produce light-emitting luciferin. In one recent study, researchers enhanced the ability to observe heart disease in laboratory mice by producing mice whose hearts emit a green-colored light with each heartbeat. They did so by deriving a green fluorescent protein from a bioluminescent species of jellyfish and modifying it. In another study, researchers transfected firefly luciferase to create mice with livers that glow during glucose production, signaling an absence of insulin, and were thus able to isolate the biochemical mechanism involved in type II diabetes.

In research conducted in China, mice were engineered with luciferase transfection to glow red, with implications for cancer research and the treatment of various diseases with gene therapy.

A fish species called zebra danio that also emits red light has been marketed in pet stores throughout the United States. The luciferase DNA for this black and white fish comes from bioluminescent sea coral.

Resource by Will

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