This is an audio version of the Wikipedia Article:
https://en.wikipedia.org/wiki/Genetic_engineering_techniques


00:03:09 1 History
00:06:48 2 Choosing target genes
00:10:27 3 Gene manipulation
00:11:07 3.1 Extraction from cells
00:12:16 3.2 Gene isolation
00:15:10 3.3 Modification
00:16:15 4 Inserting DNA into the host genome
00:17:06 4.1 Transformation
00:21:17 4.2 Transfection
00:23:56 4.3 Transduction
00:24:59 4.4 Regeneration
00:26:49 4.5 Confirmation
00:28:03 5 Gene targeting
00:31:52 5.1 Meganucleases and Zinc finger nucleases
00:33:40 5.2 TALEN and CRISPR
00:36:35 6 See also



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SUMMARY
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Genetic engineering can be accomplished using multiple techniques. There are a number of steps that are followed before a genetically modified organism (GMO) is created. Genetic engineers must first choose what gene they wish to insert, modify or delete. The gene must then be isolated and incorporated, along with other genetic elements, into a suitable vector. This vector is then used to insert the gene into the host organism, creating the GMO.
The ability to genetically engineer organisms is built on years of research and discovery on how genes function and how we can manipulate them.Humans have been manipulating genetics since early domestication attempts around 12,000 BC. Following the discovery of genes by Gregor Mendel and the proof that they were involved in inheritance tools were developed that allowed their direct manipulation. Important advances included the discovery of restriction enzymes and DNA ligases and the development of polymerase chain reaction and sequencing.
This allowed the gene of interest to be isolated and then incorporated into a vector. Often a promoter and terminator region was added as well as a selectable marker gene. The gene may be modified further at this point to make it express more efficiently. This vector is then inserted into the host organism's genome. For animals, the gene is typically inserted into embryonic stem cells, while in plants it can be inserted into any tissue that can be cultured into a fully developed plant. Common techniques include microinjection, virus-mediated, Agrobacterium-mediated or biolistics. Further tests are carried out on the resulting organism to ensure stable integration, inheritance and expression. First generation offspring are heterozygous, requiring them to be inbred to create the homozygous pattern necessary for stable inheritance. Homozygosity must be confirmed in second generation specimens.
Traditional techniques inserted the genes randomly into the hosts genome. Advances have allowed genes to be inserted at specific locations within a genome, which reduces the unintended side effects of random insertion. Early targeting systems relied on meganucleases and zinc finger nucleases. Since 2009 more accurate and easier systems to implement have been developed. Transcription activator-like effector nucleases (TALENs) and the Cas9-guideRNA system (adapted from CRISPR) are the two most commonly used. They may potentially be useful in gene therapy and other procedures that require accurate or high through put targeting.