By Feng Zhang, Holger Puchta, James G. Thomson
Over the previous 50 years, biotechnology has been the foremost driver for expanding crop productiveness. fairly, advances in plant genetic engineering applied sciences have spread out enormous new possibilities for plant researchers and breeders to create new crop types with fascinating qualities. fresh improvement of targeted genome amendment tools, reminiscent of certain gene knock-out/knock-in and designated gene substitute, strikes genetic engineering to a different point and provides much more potentials for making improvements to crop creation. The paintings offers an outline of the newest advances on distinctive genomic engineering applied sciences in crops. themes contain recombinase and engineered nucleases-mediated specific amendment, negative/positive selection-based homologous recombination and oligo nucleotide-mediated recombination. ultimately, demanding situations and affects of the hot applied sciences on current laws for genetic amendment organisms (GMOs) should be discussed.
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Extra info for Advances in New Technology for Targeted Modification of Plant Genomes
2 Engineering Meganuclease for Precise Plant Genome Modification 35 The resulting nucleases, termed as megaTAL or compact TALEN, have shown great potential with high activity and compact architectures. Taken together, a large number of engineered meganucleases have been successfully developed for targeted genome modification in numerous organisms, including human, mouse, maize, cotton, tobacco, and rice (Arnould et al. 2007; Grizot et al. 2009; Daboussi et al. 2012; Valton et al. 2012; D’Halluin et al.
2007; Redondo et al. 2008). An example of protein–protein interface engineering is the engineered obligate heterodimeric meganuclease targeting the human gene RAG1. These mutant proteins carry only a subset of compensatory mutations (K7E/E8K and E61R/K96E) that proved sufficient to destabilize homodimer formation without any loss of heterodimer activity. Consequently, the resulting meganuclease demonstrated decreased toxicity with reduced off-site cleavage events measured by the formation of phosphorylated H2AX histone foci (γ-H2AX) (Grizot et al.
2012). The proof-of-concept experiment was set up in an Arabidopsis line that contains a pre-integrated transgene as the target locus. This target locus consists of an I-SceI site upstream of a promoterless β-glucuronidase (GUS) reporter gene. In this strategy, the donor construct, which contains a 35S promoter flanked by two I-SceI sites, was transformed and integrated into the plant genome first. The resulting transgenic line was then crossed with an I-SceI-expressing line. In the hybrid line, I-SceI created DSBs in both the target locus and the donor sequences leading to excision of the donor repair template from its integration site.
Advances in New Technology for Targeted Modification of Plant Genomes by Feng Zhang, Holger Puchta, James G. Thomson