C7 protein is an

engineered Engineering is the use of scientific principles to design and build machines, structures, and other items, including bridges, tunnels, roads, vehicles, and buildings. The discipline of engineering encompasses a broad range of more specializ ...

zinc finger protein A zinc finger is a small protein structural motif that is characterized by the coordination of one or more zinc ions (Zn2+) in order to stabilize the fold. It was originally coined to describe the finger-like appearance of a hypothesized structu ...

based on the murine ZFP,

Zif268 EGR-1 (Early growth response protein 1) also known as ZNF268 (zinc finger protein 268) or NGFI-A (nerve growth factor-induced protein A) is a protein that in humans is encoded by the ''EGR1'' gene. EGR-1 is a mammalian transcription factor. It wa ...

and discovered by Wu ''et al.'' in 1994 (published in 1995). It shares the same

zinc finger A zinc finger is a small protein structural motif that is characterized by the coordination of one or more zinc ions (Zn2+) in order to stabilize the fold. It was originally coined to describe the finger-like appearance of a hypothesized struct ...

2 and zinc finger 3 of Zif268, but differs in the sequence of finger 1. It also shares the same DNA target, 5'-GCGTGGGCG-3'. The shared sequences in single letter amino acid codes of fingers 2 and 3 are RSD-H-LTT and RAD-E-RKR (positions -1 through 6 in the alpha helix). Zinc finger 1 has the sequence KSA-D-LKR which provides a 13-fold increase in affinity to the target sequence of the entire ZFP over that of Zif268. It is used in zinc finger investigations in which the amino acid sequence of finger 2 is changed in order to determine the appropriate sequence to target a given three-nucleotide target site. A variation of C7, C7.GAT is preferred since it lacks the

aspartic acid Aspartic acid (symbol Asp or D; the ionic form is known as aspartate), is an α-amino acid that is used in the biosynthesis of proteins. Like all other amino acids, it contains an amino group and a carboxylic acid. Its α-amino group is in the pro ...

residue present in finger 3 of C7 and known to cause a phenomenon called 'target site overlap'. In this case the target site overlap is a result of the aspartic acid residue forming a hydrogen bond with the N4 of the cytosine (in the opposite strand) base-paired to the guanine in the finger 2 subsite. It can also form the same hydrogen bond with an adenine base paired to a thymine. This target site overlap would dictate that either a cytosine or adenine residue be present as the 3' nucleotide in the finger 2 subsite which is unacceptable when looking to target sequences containing another nucleotide at this position.

References

* {{Transcription factors and intracellular receptors, g2 Engineered proteins Genetics experiments Molecular genetics