Lucy Cherbas1, Jennifer Hackney, Lei Gong, Claire Salzer, Eric Mauser, Dayu Zhang and Peter Cherbas.
Tools for Targeted Genome Engineering of Established Drosophila Cell Lines.
Early online in G3.
From the abstract: "We describe an adaptation of ΦC31 integrase-mediated targeted cassette exchange for use in Drosophila
cell lines. Single copies of an attP-bounded docking platform carrying a
GFP-expression marker, with or without insulator
elements flanking the attP sites, were inserted by
P-element transformation into the Kc167 and Sg4 cell lines; each of the
resulting docking site lines carries a single
mapped copy of one of the docking platforms. ... We describe procedures for isolating cells carrying the
substitutions ... When
compared with clonal lines made by
traditional transformation methods ... targeted insertion lines
give
more uniform expression, lower basal expression and
higher induction ratios. Targeted substitution, though intricate,
affords
results that should greatly improve comparative
expression assays – a major emphasis of cell-based studies."
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