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The high sequence specificity of minor groove-binding  N -methylpyrrole- N -methylimidazole polyamides have made significant advances in cancer and disease biology, yet there have been few comprehensive reports on their off-target effects, most likely as a consequence of the lack of available tools in evaluating genomic binding, an essential aspect that has gone seriously underexplored. Compared to other  N -heterocycles, the off-target effects of these polyamides and their specificity for the DNA minor groove and primary base pair recognition require the development of new analytical methods, which are missing in the field today. This review aims to highlight the current progress in deciphering the off-target effects of these  N -heterocyclic molecules and suggests new ways that next-generating sequencing can be used in addressing off-target effects.

The Road Not Taken with Pyrrole-Imidazole Polyamides: Off-Target Effects and Genomic Binding