Viral Construct Design and Packaging

The CreOn-FlpOff and FlpOn-CreOff ExBoX viral constructs were designed using ApE (https://jorgensen.biology.utah.edu/wayned/ape/) and Vector Builder (https://en.vectorbuilder.com/) free software. The plasmids were synthesized and assembled by Vector Builder. Plasmid DNA was purified using the Qiagen Maxi Prep kit (Qiagen Cat#10023). AAVs were also packaged by Vector Builder into AAV2/9. pENN.AAV.CamKII 0.4. Cre. SV40 (AAV5) (deposited by James M. Wilson; Addgene #105558; RRID:Addgene_105558), pAAV-CAG-TdTomato (AAV5) (deposited by Edward Boyden; Addgene #59462; RRID:Addgene_59462) and pCAG-FLEX-EGFP-WPRE (AAV9) (deposited by Hongkui Zeng; Addgene #51502;; RRID:Addgene_51502) were obtained from Addgene (Oh et al., 2014 (link)). rAAV5/AAV-EF1α-FlpO-WPRE and rAAV9/CAG-GFP were obtained from the UNC GTC Vector Core. pENN.AAV.hSyn.Cre.hGH was Addgene #105555 (RRID:Addgene_105555) deposited by James M. Wilson. All the ExBoX constructs designed for this study, along with the maps and sequences, have been deposited in Addgene. Addgene IDs of ExBoX plasmids can be found in Table S2.

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Ubina T., Vahedi-Hunter T., Agnew-Svoboda W., Wong W., Gupta A., Santhakumar V, & Riccomagno M.M. (2021). ExBoX – a simple Boolean exclusion strategy to drive expression in neurons. Journal of Cell Science, 134(20), jcs257212.

Publication 2021

Maxi Prep kit pAAV-CAG-TdTomato pCAG-FLEX-EGFP-WPRE

Camkii Creon Maps Plasmids Sv40 Tdtomato Vector

Corresponding Organization :

Other organizations : University of California, Riverside

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Variable analysis

independent variables

CreOn-FlpOff and FlpOn-CreOff ExBoX viral constructs

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive controls: pENN.AAV.CamKII 0.4. Cre. SV40 (AAV5), pAAV-CAG-TdTomato (AAV5), pCAG-FLEX-EGFP-WPRE (AAV9)
Negative controls: Not explicitly mentioned

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