2017
DOI: 10.1002/cbic.201700458
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Nested PUF Proteins: Extending Target RNA Elements for Gene Regulation

Abstract: RNA-binding proteins recognizing unique sequences within large transcriptomes serve as a powerful tool to control RNA metabolism. Pumilio and fem-3 mRNA-binding factor (PUF) proteins are considered good candidates for such tools, because they are typically composed of eight highly homologous repeat segments and can be designed to recognize arbitrary 8 nt RNA sequences. However, a specific 8 nt RNA sequence is found at multiple sites in various RNAs in the transcriptome, making it difficult to specifically targ… Show more

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Cited by 8 publications
(18 citation statements)
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“…Firefly luciferase activity (Fluc) was normalized to Renilla luciferase activity (Rluc), and then relative luciferase activities (RLAs), compared to control samples, were calculated (Equation ( 1 1, Figure S2b). Although the K d values were somewhat different between PUFs, they were within the reported range of engineered PUF proteins (10 -10 -10 -8 M) [23,31]. Importantly, all PUF (A)-(D) proteins showed high specificity to their corresponding RNA sequences (Figure S2a).…”
Section: Luciferase Reporter Assaymentioning
confidence: 54%
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“…Firefly luciferase activity (Fluc) was normalized to Renilla luciferase activity (Rluc), and then relative luciferase activities (RLAs), compared to control samples, were calculated (Equation ( 1 1, Figure S2b). Although the K d values were somewhat different between PUFs, they were within the reported range of engineered PUF proteins (10 -10 -10 -8 M) [23,31]. Importantly, all PUF (A)-(D) proteins showed high specificity to their corresponding RNA sequences (Figure S2a).…”
Section: Luciferase Reporter Assaymentioning
confidence: 54%
“…PUF proteins were expressed in E. coli BL21 (DE3) cells (Nippon gene) and purified as described before [23]. Briefly, protein expression was induced by adding 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG), and E. coli cells were incubated at 18 • C overnight.…”
Section: Protein Expression and Purificationmentioning
confidence: 99%
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