Double-strand break repair pathways differentially affect processing and transduction by dual AAV vectors

Recombinant adeno-associated viral vectors (rAAV) are a powerful tool for gene delivery but have a limited DNA carrying capacity. Efforts to expand this genetic payload have focused on engineering the vector components, such as dual trans-splicing vectors which double the delivery size by exploiting the natural concatenation of rAAV genomes in host nuclei. We hypothesized that inefficient dual vector transduction could be improved by modulating host factors which affect concatenation. Since factors mediating concatenation are not well defined, we performed a genome-wide screen to identify host cell regulators. We discovered that Homologous Recombination (HR) is inhibitory to dual vector transduction. We demonstrate that depletion or inhibition of HR factors BRCA1 and Rad51 significantly increase reconstitution of a large split transgene by increasing both concatenation and expression from rAAVs. Our results define new roles for DNA damage repair in rAAV transduction and highlight the po..., A Cas9 KO screen was conducted to recover regulators of dual-vector AAVS transduction in U2OS cells. This code operates on outputs from the hypergeometric analysis in the Broad Institute's GPP Web Portal and pathway analysis from Reactome. It primarily tabulates and plots these results., , # Data in support of \"Double-Strand Break Repair Pathways Differentially Affect Processing and Transduction by Dual AAV Vectors\"

[https://doi.org/10.5061/dryad.7h44j1053](https://doi.org/10.5061/dryad.7h44j1053)

### Files

\"**hypergeometric.txt**\" is the output of the Broad Institute's hypergeometric screen analysis.

Headers:

* \"Gene Symbol\": alphanumeric designation for the target of the perturbations
* \"Average LFC\": mean of log-fold change in guide abundance 
* \"Average -log(p-values)\": mean of negative log10 p-values associated with guide log-fold change
* \"Number of perturbations\": number of guide sequences recovered corresponding to this gene
* \"Perturbations\": guide sequences, same number as \"Number of perturbations\",  separated by semicolons
* \"Individual LFCs\": log-fold changes in guide abundances separated by semicolons. These were averaged to yield \"Average LFC\".
* \"Ascending ranks\": perturbation-wise log-fold change in guide abundances in increasing order separated by se...