journal club 11/25/13

Jiang He

Fluorescent proteins as a scaffold for cell-specific gene manipulation

Background

  • Lots of GFP lines
  • not a lot of Cre / lox or Gal4 lines (etc) for manipulating gene expression.
  • camilid VHH antigen binding domain (‘nanobody’).
  • Previous work: different nanobodies against GFP change it’s fluorescent intensity (‘enhancers’ and ‘minimizers’)

This paper

  • screen for GFP nanobodies
  • recruit VP16 to target genes in a GFP dependent manner.
  • why not genetically encode, reporter gene and nano-bodies, then cross this to a GFP mouse line.
  • Concern: low level GFP still respond
  • GFP lines not very specific — have a high expressing cell type and a low expressing cell type.
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