Solid-phase synthesis of highly repetitive chromatin assembly templates

- Solid-phase DNA ligation method to generate unique chromatin model system templates.
- Optimized to produce either desired template length or diversity of template sizes.
- DNA ligation monomers include both single and multi-nucleosome binding sites.
- Synthesized templates assemble into chromatin model systems.

DNA templates for assembling chromatin model systems typically consist of numerous repeats of nucleosome positioning sequences, making their synthesis challenging. Here we describe a solid-phase strategy for generating such templates using sequential enzymatic ligation of DNA monomers. Using single nucleosome site monomers, we can either generate a twelve-nucleosome site target, or systematically access intermediate-sized templates. Using twelve nucleosome positioning site monomers, longer templates can be generated. Our synthesized templates assemble into well-defined chromatin model systems, demonstrating the utility of our solid-phase approach. Moreover, our strategy should be more widely applicable to generating other DNAs containing highly repetitive DNA sequences.