Insertion of PATC-rich C. elegans introns into synthetic transgenes by golden-gate-based cloning

Abstract

Transgenes are particularly prone to epigenetic silencing in the C. elegans germline. Here, we describe a protocol to insert introns containing a class of non-coding DNA named Periodic An/Tn Clusters (PATCs) into synthetic transgenes. PATCs can protect transgenes from position-dependent silencing (Position Effect Variegation, PEV) and from silencing in simple extra-chromosomal arrays. Using a set of simple design rules, it is possible to routinely insert up to three PATC-rich introns into a synthetic transgene in a single reaction.