Rapid Assembly and Cloning of Zinc Finger Proteins with Multiple Finger Modules

摘要

The C2H2-type zinc finger (ZF) is one of the most abundant structural motifs among eukaryotes, exceeding over 3% of the whole human genes. Primarily, the ZF plays an important role in recognizing nucleic acid targets including DNA and RNA in a sequence-specific fashion. In-depth studies of the ZF domain from the transcription factor IIIA have provided vital informations about the ZF on the nucleic acid recognition principle. A ZF recognizes DNA principally by one-to-one interaction between amino acids and DNA bases. Amino acids at helical positions -1, +3, and +6 of the recognition helix form specific interactions with three consecutive DNA bases on one strand of the DNA, although there is an additional interaction from the position +2 to the other strand. Many studies demonstrate that ZF can be modularly engineered to alter their sequence-specificity. In particular, the phage-display is a powerful technique to screen and select novel sequence-specific ZF peptides that bind to various DNA sequences. Accumulating data in engineered ZFs demonstrates that almost any DNA sequences can be recognized by one-to-one interaction between ZFs and triplet bases. Accordingly, the ZFs are an ideal frame for designing new DNA binding proteins to recognize any given sequence of DNA.