By Feng Zhang, Holger Puchta, James G. Thomson
Over the previous 50 years, biotechnology has been the most important driver for expanding crop productiveness. really, advances in plant genetic engineering applied sciences have spread out great new possibilities for plant researchers and breeders to create new crop forms with fascinating features. fresh improvement of exact genome amendment equipment, corresponding to designated gene knock-out/knock-in and detailed gene alternative, strikes genetic engineering to a different point and gives much more potentials for bettering crop construction. The paintings presents an outline of the most recent advances on detailed genomic engineering applied sciences in crops. subject matters contain recombinase and engineered nucleases-mediated designated 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 info for Advances in New Technology for Targeted Modification of Plant Genomes
Use of meganucleases in precise genome modification requires the ability to engineer nucleases with new sequence specificity. One challenge of meganuclease engineering is that cleavage and DNA-binding domains overlap (Stoddard 2011). When an amino acid sequence is altered to achieve new DNA sequence specificity, the catalytic activity of the enzyme is often compromised. Despite these difficulties, advances in high-throughput screening and protein modeling have made meganucleases easier to engineer and more accessible.
2003). 2 Principle of the Two-Step Semi-rational Approach In this method, the meganuclease I-CreI was chosen as the molecular scaffold to produce novel meganucleases. As indicated in Fig. 2, I-CreI works as a homodimer on a pseudo-palindromic target. The DNA–protein interaction is accomplished via a region involving 40 amino acids. As previously reported, each monomer can be divided into sub-domains. For instance, we described two sub-domains referred to as distal (10NNN) and proximal (5NNN) to the catalytic center (Fig.
2008). The mutations of K7E/E8K, E61R/K96E, and R51D/D137R in homodimeric I-CreI meganucleases have led to the generation of obligate I-CreI-derived heterodimeric nucleases (Grizot et al. 2009). In addition, the mutation of G19S has been shown to have a major impact on decreasing homodimer activity, with no decrease in heterodimer activity (Arnould 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.
Advances in New Technology for Targeted Modification of Plant Genomes by Feng Zhang, Holger Puchta, James G. Thomson