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FROM GFP TO HALO
Protein Labeling Inside a Cell
Xi Wu
The Jackson Laboratory
University of Maine
Stereocilia bundle in mouse cochlea
hair cell.
Neuron cells in Drosophila larvae imaginal
discs.
Crystal structure of GFP. GFP folds into a 11 beta-sheet barrel; a
chromophore structure in within the barrel.
Tracking protein of interest
Crivat et al. (2012) Trends in Biotech. 30: 8-16
Johnsson et al. (2012) NEB expressions Fall 2008, Vol. 3.3
KpnI
Forward primer
GNAI2 gene fused
with EGFP
Caggs
Gnai2
EGFP
NotI
Reverse primer
PCR clone
KpnI
Forward primer
GNAI2 gene fused
with SNAP/Halo tag
Caggs
SNAP
or
Halo
Gnai2
NotI
Reverse primer
Cochlea Electroporation
CRISPR/Cas9 Genome Editing
Figure courtesy of Delivery by Mirus.
Pennisi et al., Science, 2013, 341:833-836.
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