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• W2149259570 abstract "The ability to engineer biological circuits that process and respond to complex cellular signals has the potential to impact many areas of biology and medicine. Transcriptional activator-like effectors (TALEs) have emerged as an attractive component for engineering these circuits, as TALEs can be designed de novo to target a given DNA sequence. Currently, however, the use of TALEs is limited by degeneracy in the site-specific manner by which they recognize DNA. Here, we propose an algorithm to computationally address this problem. We apply our algorithm to design 180 TALEs targeting 20 bp cognate binding sites that are at least 3 nt mismatches away from all 20 bp sequences in putative 2 kb human promoter regions. We generated eight of these synthetic TALE activators and showed that each is able to activate transcription from a targeted reporter. Importantly, we show that these proteins do not activate synthetic reporters containing mismatches similar to those present in the genome nor a set of endogenous genes predicted to be the most likely targets in vivo. Finally, we generated and characterized TALE repressors comprised of our orthogonal DNA binding domains and further combined them with shRNAs to accomplish near complete repression of target gene expression." @default.
• W2149259570 created "2016-06-24" @default.
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• W2149259570 date "2012-05-11" @default.
• W2149259570 modified "2023-09-29" @default.
• W2149259570 title "Engineering synthetic TAL effectors with orthogonal target sites" @default.
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• W2149259570 doi "https://doi.org/10.1093/nar/gks404" @default.
• W2149259570 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3424557" @default.
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• W2149259570 hasPublicationYear "2012" @default.
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