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DNA constructs

VP Veselina Petrova
CP Craig S. Pearson
JC Jared Ching
JT James R. Tribble
AS Andrea G. Solano
YY Yunfei Yang
FL Fiona M. Love
RW Robert J. Watt
AO Andrew Osborne
ER Evan Reid
PW Pete A. Williams
KM Keith R. Martin
HG Herbert M. Geller
RE Richard Eva
JF James W. Fawcett
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Human Protrudin constructs (in pmCherry-C1 and pEGFP-C1)48, CMV-integrin-alpha9-GFP29 and CMV-Rab11 -GFP19,29 were used. The CMV promoter in all constructs was replaced by a human synapsin (Syn) promoter by Gibson assembly cloning. The viral vector plasmid backbones (AAV2-sCAG-GFP) were a kind donation by Prof. Joost Verhaagen, The Netherlands Institute for Neuroscience. Protrudin-GFP was cloned from pEGFP-C1 plasmid into viral vector plasmids using Gibson cloning. Site-direct mutagenesis was performed in order to create the Protrudin active phosphomimetic form (QuikChange II Site-Directed Mutagenesis Kit, Agilent Technologies). All primers used for Protrudin cloning are described in Table S1. All constructs were verified by DNA sequencing.

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