Figure 2.
Engineering of opto-kinesin.(A) Design of opto-kinesin. A blue light–sensitive kinesin was engineered by replacing the neck coil (aa 366–383) of KIF1A(1–383)-GFP-SSPB with the blue light–sensitive homodimerization module VVD52C, generating KIF1A(1–365)-VVD52C-GFP-SSPB (opto-kinesin). (B) Schematic representation of the VVD52C and VVDfast modules. (C) Live-cell imaging of KIF1A(1–365)-VVD52C-mCherry-SSPB(micro) in U2OS cells before and after 10 min of illumination (see also Video 1). Scale bar is 10 µm. (D–F) Quantification of experiments as in C depicting normalized peripheral intensity of the two opto-kinesins relative to preillumination state following continuous (D) or pulsed (E and F) illumination (470 nm; blue box). Graphs in D and E represent mean and SEM of at least four cells.

Engineering of opto-kinesin. (A) Design of opto-kinesin. A blue light–sensitive kinesin was engineered by replacing the neck coil (aa 366–383) of KIF1A(1–383)-GFP-SSPB with the blue light–sensitive homodimerization module VVD52C, generating KIF1A(1–365)-VVD52C-GFP-SSPB (opto-kinesin). (B) Schematic representation of the VVD52C and VVDfast modules. (C) Live-cell imaging of KIF1A(1–365)-VVD52C-mCherry-SSPB(micro) in U2OS cells before and after 10 min of illumination (see also Video 1). Scale bar is 10 µm. (D–F) Quantification of experiments as in C depicting normalized peripheral intensity of the two opto-kinesins relative to preillumination state following continuous (D) or pulsed (E and F) illumination (470 nm; blue box). Graphs in D and E represent mean and SEM of at least four cells.

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