Abstract
The emergence of genome manipulation methods promises a real revolution in biotechnology and genetic engineering. Targeted editing of the genomes of living organisms not only permits investigations into the understanding of the fundamental basis of biological systems but also allows addressing a wide range of goals towards improving productivity and quality of crops. This includes the creation of plants with valuable compositional properties and with traits that confer resistance to various biotic and abiotic stresses. During the past few years, several novel genome editing systems have been developed; these include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats/Cas9 (CRISPR/Cas9). These exciting new methods, briefly reviewed herein, have proved themselves as effective and reliable tools for the genetic improvement of plants.
Highlights
Since the advent of recombinant DNA technology in Paul Berg’s laboratory [1] in 1972, genetic engineering has come a long way and achieved enormous success
In 1996, for the first time, it was shown that protein domains such as “zinc fingers” coupled with FokI endonuclease domains act as site-specific nucleases (zinc finger nucleases (ZFNs)), which cleave the DNA in vitro in strictly defined regions [3]
We briefly described the mechanisms of different genome editing systems and their use for crop improvement and highlighted the multiple advantages and applications of engineered nucleases as well as biosafety and regulatory aspects of plants generated using engineered nuclease based technologies
Summary
Since the advent of recombinant DNA technology in Paul Berg’s laboratory [1] in 1972, genetic engineering has come a long way and achieved enormous success. Several decades-long investigations in molecular genetics and biochemistry of bacteria and viruses have allowed researchers to develop new methods of manipulating DNA through creation of various vector systems and tools for their delivery into the cell. All of these developments allow successful creation of transgenic microorganisms and genetically modified higher organisms including various plant and crop species. In 1996, for the first time, it was shown that protein domains such as “zinc fingers” coupled with FokI endonuclease domains act as site-specific nucleases (zinc finger nucleases (ZFNs)), which cleave the DNA in vitro in strictly defined regions [3] Such a chimeric protein has a modular structure, because each of the “zinc finger” domains recognizes one triplet of nucleotides. We briefly described the mechanisms of different genome editing systems and their use for crop improvement and highlighted the multiple advantages and applications of engineered nucleases as well as biosafety and regulatory aspects of plants generated using engineered nuclease based technologies
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