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Expanding RNA binding specificity and affinity of engineered PUF domains.

Nucleic Acids Res.. 2018; 
Zhao Yang-Yang,Mao Miao-Wei,Zhang Wen-Jing,Wang Jue,Li Hai-Tao,Yang Yi,Wang Zefeng,Wu Jia
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Nucleic Acid Purification & Analysis and heated at 95◦C for 10 min. Then the mixtures were separated by 4–20% SDS-PAGE Gel (Genscript Get A Quote

摘要

Specific manipulation of RNA is necessary for the research in biotechnology and medicine. The RNA-binding domains of Pumilio/fem-3 mRNA binding factors (PUF domains) are programmable RNA binding scaffolds used to engineer artificial proteins that specifically modulate RNAs. However, the native PUF domains generally recognize 8-nt RNAs, limiting their applications. Here, we modify the PUF domain of human Pumilio1 to engineer PUFs that recognize RNA targets of different length. The engineered PUFs bind to their RNA targets specifically and PUFs with more repeats have higher binding affinity than the canonical eight-repeat domains; however, the binding affinity reaches the peak at those with 9 and 10 repea... More

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