AAV vectors were produced by the triple transfection method using HEK293T cells [31 (link),32 (link)]. The shuttle plasmids pAAV[shRNA]-EGFP-U6 > mZnf219[shRNA#1] or [shRNA scramble] (VectorBuilder) were packaged into AAV9 capsids using the helper plasmids pAdDF6 and plasmid pAAV9 (providing the rep and cap viral genes) (PennVector). All plasmids were co-transfected into HEK293T cells using linear polyethylenimine (MW 25,000). The cells were seeded in Hyperflasks (Corning) at 1.2 × 108 cells per flask the day before transfection.
Transfection and viral particle collection were performed as previously described [33 (link)]. Sequences of sh-RNAs are included inTable S4 .
Transfection and viral particle collection were performed as previously described [33 (link)]. Sequences of sh-RNAs are included in
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