Tuesday, November 24, 2015

Drosophila cell team at the DGRC reports integrase-mediated cassette exchange approach to production of clonal transgenic fly cell lines

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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