Figure 5.

Effect of sequence variation in the DNA spacer library on Tev-mTALEN activity. (A) Activity was screened on a library of substrates with random spacer sequences, with TP_15N represented in the schematic at top relative to the wild-type T4 substrate, TP15. Shown below is a representative example of substrates tested in a 96-well microtiter plate assay in which the individual wells are colored according β-galactosidase activity (in Miller units). The red rectangles at the top right indicate the positive and negative controls. Yellow rectangles indicate substrates on which activity was greater than or within 2 SDs of the wild-type control, averaged over three technical replicates. (B) Logos plot of information content (in bits) (Schneider and Stephens 1990) per position for 49 substrates that showed high activity (cleavers) and 62 substrates with background activity (noncleavers), respectively.