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Targeted DNA Methylation Using an Artificially Bisected M.HhaI Fused to Zinc Fingers

Figure 4

Molecular modeling explains how an increase in target spacing can reduced the required protein linker length.

(A) HS1 zinc fingers are bound to DNA with a target site spacing of Z = 0,1,2, or 3. Note that at Z = 3, the zinc finger is actually closer to the bisection point of the N-termini than at Z = 0. (B) A model demonstrating that a five amino acid linker is sufficient to connect zinc finger HS1 bound at Z = 3. In contrast, a longer amino acid linker is required to circumvent the DNA backbone at Z = 0. This model provides an explanation for the pattern of target site methylation observed in Figure 3D.

Figure 4