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.
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Posted 15 Feb, 2021
Posted 15 Feb, 2021
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.
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