Journal: Cell Reports Methods

Article Title: A versatile viral toolkit for functional discovery in the nervous system

doi: 10.1016/j.crmeth.2022.100225

Figure Lengend Snippet: Constitutive elements analysis of Addgene most commonly used AAV vectors Number of occurrences of the indicated elements among Addgene’s top plasmids sorted by size (base pairs, bp). (A) Backbones. The dotted line shows the 4.7 kb payload limit. (B) Promoters. WPRE and polyA sequences. (C) Distribution of backbones’ sizes for recombinase-inducible plasmids from Addgene. Each row represents specific recombinase dependencies as described on the left y axis, and its corresponding VTK constructs are represented on the right y axis. (D) Number of occurrences of backbones in direct comparison to Cre-ON conformation. In (A–D), the colored dots on the x axis indicate the size of the corresponding elements used in VTK vectors. Green dots for common elements to all VTKs (WPRE and polyA), purple dots for VTKS1–6, and yellow dots for VTKD1–6. See also Figure S1 and Table S1 .

Article Snippet: VTKS/D constructs containing the promoter and inducible sites were synthesized (see ) and cloned using ApaI/AscI into a backbone containing ITRs, WPRE and bGHp polyA (from Addgene #52473, a generous gift from Ian Wickersham) ( ).

Techniques: Construct, Comparison

Journal: Cell Reports Methods

Article Title: A versatile viral toolkit for functional discovery in the nervous system

doi: 10.1016/j.crmeth.2022.100225

Figure Lengend Snippet: Optimization of a viral toolkit (VTK) for direct and combinatorial transgene expression Diagram representing each VTK plasmid. The size between ITRs and the corresponding Boolean strategy are displayed below the relevant plasmid backbone. Top row: VTKS1–6. Bottom row: VTKD1–6. MCS, multiple cloning site; pA: polyA; LoxP, Lox2272 are Cre-dependent sites; FRT, F5 are Flp-dependent sites. See also Figure S2 and Table S2 .

Article Snippet: VTKS/D constructs containing the promoter and inducible sites were synthesized (see ) and cloned using ApaI/AscI into a backbone containing ITRs, WPRE and bGHp polyA (from Addgene #52473, a generous gift from Ian Wickersham) ( ).

Techniques: Expressing, Plasmid Preparation, Cloning