Download e-book for kindle: Herbal Medicine for Human Health by R.R. Chaudhury

By R.R. Chaudhury

ISBN-10: 9290221623

ISBN-13: 9789290221623

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TALEs are composed of N-terminal secretion and translocation signals, a central domain with DNA binding capability and a C-terminal acidic activator domain coupled to nuclear localization signals that enable translocation into nucleus [2] (Fig. 1). Protein engineering strategies have been especially focused on the identification of N- or C-terminal truncations aimed at creating artificial TALE-based DNA binding domains that combine minimal size with efficient DNA binding activity. In the next paragraphs we will describe and summarize the development of different TALE scaffolds that have been engineered as versatile carriers for various effector domains.

The first ZFNs for a genomic target displayed significant toxicity when expressed at high levels [38], due to promiscuous binding and cleavage [39], and this issue has continued to trouble many new designs. Increasing the number of fingers in each monomer is one approach to ameliorating this effect, but that is not always sufficient [40, 41]. The toxicity is frequently a property of one ZFN within a pair, and it appears to be due to inadequate specificity, leading to homodimerization and cleavage at unintended, off-target sites [39].

The latter approach seems to work because of the high intrinsic positive charge on the ZFNs, and it is applicable to a range of cell types, albeit with variable efficiencies. Long, doublestranded donor DNAs can be introduced on plasmid or viral vectors, while short, single-stranded donors are usually simply added to the medium [54, 55, 58, 59]. For situations in which manipulation of whole organisms is the goal, genome alterations in the germ line must be achieved. The cells in which the germ line is most accessible are typically in the very early embryo.

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Herbal Medicine for Human Health by R.R. Chaudhury


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