By Feng Zhang, Holger Puchta, James G. Thomson
Over the previous 50 years, biotechnology has been the key motive force for expanding crop productiveness. relatively, advances in plant genetic engineering applied sciences have spread out substantial new possibilities for plant researchers and breeders to create new crop kinds with fascinating characteristics. contemporary improvement of exact genome amendment equipment, corresponding to detailed gene knock-out/knock-in and specified gene substitute, strikes genetic engineering to a different point and provides much more potentials for bettering crop construction. The paintings offers an summary of the newest advances on specific genomic engineering applied sciences in crops. subject matters contain recombinase and engineered nucleases-mediated special amendment, negative/positive selection-based homologous recombination and oligo nucleotide-mediated recombination. eventually, demanding situations and affects of the recent applied sciences on current laws for genetic amendment organisms (GMOs) can be discussed.
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Extra resources for Advances in New Technology for Targeted Modification of Plant Genomes
Nature 275(5678):336–338 Certo MT, Gwiazda KS, Kuhar R et al (2012) Coupling endonucleases with DNA end-processing enzymes to drive gene disruption. Nat Methods 9(10):973–975 Chames P, Epinat JC, Guillier S, Patin A, Lacroix E, Paques F (2005) In vivo selection of engineered homing endonucleases using double-strand break induced homologous recombination. Nucleic Acids Res 33(20):e178 Chen Z, Zhao H (2005) A highly sensitive selection method for directed evolution of homing endonucleases. Nucleic Acids Res 33(18):e154 Chevalier BS, Stoddard BL (2001) Homing endonucleases: structural and functional insight into the catalysts of intron/intein mobility.
The HR events can then be readily identified by GUS staining. By screening F2 seedlings from the hybrid line, approximately 1 % of plants were identified and confirmed as the recombinant lines with precise HR events. The high efficiency observed in this study makes this strategy very attractive for gene targeting in plant species that don’t have efficient transformation systems. In addition to facilitating targeted gene insertions in plant and animal species, engineered meganucleases have also been employed to create targeted knockout mutations in genes of interest.
2014; Boissel et al. 2014). Because all the meganucleases fused in megaTALs are monomeric endonucleases, the new hybrid nuclease does not require dimerization to cleave target sites. Moreover, DNA binding specificity of megaTALs is determined by both the TALE and meganuclease, reducing the likelihood that more than one target site would be present in the targeted genome. Thus, the fusion nucleases could offer higher specificity than regular meganucleases. In a recent research study, Boissel et al.