Data for: Growth cone advance requires EB1 as revealed by genomic replacement with a light-sensitive variant
Creators
- 1. University of California, San Francisco
- 2. Erasmus University Medical Center
Description
A challenge in analyzing dynamic intracellular cell biological processes is the dearth of methodologies that are sufficiently fast and specific to perturb intracellular protein activities. We previously developed a light-sensitive variant of the microtubule plus end-tracking protein EB1 by inserting a blue light-controlled protein dimerization module between functional domains. Here, we describe an advanced method to replace endogenous EB1 with this light-sensitive variant in a single genome editing step, thereby enabling this approach in human induced pluripotent stem cells (hiPSCs) and hiPSC-derived neurons. We demonstrate that acute and local optogenetic EB1 inactivation in developing cortical neurons induces microtubule depolymerization in the growth cone periphery and subsequent neurite retraction. In addition, advancing growth cones are repelled from areas of blue light exposure. These phenotypes were independent of the neuronal EB1 homolog EB3, revealing a direct dynamic role of EB1-mediated microtubule plus end interactions in neuron morphogenesis and neurite guidance.
Notes
Raw microscopy data are provided as Nikon ND2 files that are native to Nikon Elements microscopy software. ND2 files can also be opened with ImageJ/Fiji with the Bioformats plugin (Note: metadata may not be imported correctly into ImageJ), or with a freely available Nikon Elements Viewer.
Data analysis on which manuscript figures are based are provided as Microsoft Excel spreadsheets.
Raw sequence data are provided in AB1 and SEQ file formats.
Funding provided by: National Cancer Institute
Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000054
Award Number: R21 CA224194
Funding provided by: National Institute of Neurological Disorders and Stroke
Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000065
Award Number: R01 NS107480
Funding provided by: National Institutes of Health
Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000002
Award Number: S10 RR026758
Funding provided by: National Institute of General Medical Sciences
Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000057
Award Number: S10 OD028611
Funding provided by: National Institutes of Health
Crossref Funder Registry ID: http://dx.doi.org/10.13039/100000002
Award Number: S10 OD028611
Files
Data_Analysis.zip
Additional details
Related works
- Is cited by
- 10.1101/2022.09.22.509085 (DOI)
- 10.7554/elife.84143 (DOI)