Genome-Scale Mapping of Human LINE-1 ORF2p Endonuclease Activity

LINE-1 is the only active protein-coding transposon in the human genome. Its insertions favor a 5′-TTTTT↓AA consensus motif, but it has been unclear whether that preference comes from where the ORF2p endonuclease nicks DNA or from the later reverse transcription step. This study used electronic genome mapping (EGM) to measure endonuclease nicking directly at genome scale.

Purified ORF2p endonuclease and an inactive mutant control were applied to Lambda, E. coli, and human DNA and analyzed on the OhmX™ Platform, with analysis adjusted so that low-level nicking could be detected. The poster shows how nicking at the consensus motif responds to endonuclease dose and what the remaining nick sites reveal about where the endonuclease cuts.

Explore how genome-scale nick mapping with EGM can help characterize DNA nicking and modification in your research.