Library transgenesis in zebrafish through delayed site-specific mosaic integration for in vivo pooled screening of transgenes

This dataset is from a study developing a zebrafish library transgenesis method using delayed site-specific integration to achieve single-transgene-per-cell mosaicism with high library diversity. The raw imaging data includes confocal Z-stacks (.nd2 files) of zebrafish larvae expressing a 2-component library of fluorescent proteins (GFP and mScarlet) in neurons, captured across multiple brain regions to assess transgene mutual exclusivity in mosaic animals. The raw sequencing data (.fastq files) include Illumina amplicon reads generated from genomic DNA of 12 mosaic larvae with integrated libraries of barcoded GFP constructs (6978-R2), as well as the injected source barcode library (7462-R1). Larvae were injected with a complex library of barcoded plasmids, enabling quantification of independent integration events.
, , # Library transgenesis in zebrafish through delayed site-specific mosaic integration for in vivo pooled screening of transgenes

Dataset DOI: [10.5061/dryad.d2547d8h0](https://doi.org/10.5061/dryad.d2547d8h0)

## Description of the data and file structure

This dataset contains confocal microscopy images and amplicon sequencing data characterizing a zebrafish library transgenesis method using delayed site-specific integration. This dataset includes all the raw imaging data associated with Fig. 2, S1, S2, and Table S1, and all the raw Illumina sequencing data associated with Fig. 3, S3, S4, S5, S,6 and Tables S2, S3.

### Files and variables

### GFP-mScarlet Fluorescent Library Imaging

**Format:** `.nd2` (Nikon raw imaging format, confocal hyperstacks)**.**

**Experimental setup:** pIGLET embryos were injected wita h 50:50 mix of GFP-CAAX and mScarlet-CAAX plasmids. Larvae imaged at 3 dpf or 5 dpf, live or fixed in 4% PFA, mounted in 1% agarose. Spinning disk confocal microscopy (Yoko...,