Gene amplification is a procedure in which a certain gene or DNA sequence is replicated many times in a process called DNA replication.
In separation and detection, DNA and mRNA are isolated from cells and then detected simply by the isolation. Cell cultures are also grown to provide a constant supply of cells ready for isolation.
First, laboratories use a normal cellular modification of mRNA that adds up to 200 adenine nucleotides to the end of the molecule (poly(A) tail). Once this has been added, the cell is ruptured and its cell contents are exposed to synthetic beads that are coated with thymine string nucleotides. Because Adenine and Thymine pair together in DNA, the poly(A) tail and synthetic beads are attracted to one another, and once they bind in this process the cell components can be washed away without removing the mRNA. Once the mRNA has been isolated, reverse transcriptase is employed to convert it to single-stranded DNA, from which a stable double-stranded DNA is produced using DNA polymerase. Complementary DNA (cDNA) is much more stable than mRNA and so, once the double-stranded DNA has been produced it represents the expressed DNA sequence scientists look for.
This technique is used to identify which genes or genetic mutations produce a certain phenotype. A mutagen is very often used to accelerate this process. Once mutants have been isolated, the mutated genes can be molecularly identified.
Forward saturation genetics is a method for treating organisms with a mutagen, then screens the organism's offspring for particular phenotypes. This type of genetic screening is used to find and identify all the genes involved in a trait.
A mutation in a gene can cause encoded proteins and the cells that rely on those proteins to malfunction. Conditions related to gene mutations are called genetic disorders. However, altering a patient's genes can sometimes be used to treat or cure a disease as well. Gene therapy can be used to replace a mutated gene with the correct copy of the gene, to inactivate or knockout the expression of a malfunctioning gene, or to introduce a foreign gene to the body to help fight disease. Major diseases that can be treated with gene therapy include viral infections, cancers, and inherited disorders, including immune system disorders.
Gene therapy delivers a copy of the missing, mutated, or desired gene via a modified virus or vector to the patient's target cells so that a functional form of the protein can then be produced and incorporated into the body. These vectors are often siRNA. Treatment can be either in vivo or ex vivo. The therapy has to be repeated several times for the infected patient to continually be relieved, as repeated cell division and cell death slowly randomizes the body's ratio of functional-to-mutant genes. Gene therapy is an appealing alternative to some drug-based approaches, because gene therapy repairs the underlying genetic defect using the patients own cells with minimal side effects. Gene therapies are still in development and mostly used in research settings. All experiments and products are controlled by the U.S. FDA and the NIH.