Published December 13, 2022 | Version v1
Dataset Open

Raw Sanger sequences of the tyr fragment from crispant and control zebrafish embryos

Authors/Creators

  • 1. Bambino Gesù Children's Hospital

Description

Thanks to the amenability to genetic modifications, the minimal or non-invasive phenotyping possibilities in early embryos and larvae and the presence of a high percentage of genes orthologous to humans, zebrafish is a successful alternative vertebrate to validate new disease genes and variants and to study mechanisms underlying human diseases. We developed an optimized method allowing early and gentle genotype detection to reduce further possible suffering associated with the generation of genetically modified zebrafish and valid to strategize the employment of "surplus" animals (e.g., those needed solely for line generation or without the desired genotype). The method is based on minimally-invasive tissue (fin) scratching (FS) to obtain genomic DNA material and perform genotyping in early zebrafish embryos. We showcase the method's usefulness for various genotyping needs, including screening both F0 and stable CRISPR/Cas9 lines and obtaining genomic material compatible with Sanger sequencing. Here we provide the raw sequences obtained from FS-derived genomic DNA of single mutant F0 embryos carrying mutations in the tyr gene generated with Base editor (BE) CRISPR/Cas9 technology. We compared the sequencing outcome of FS- vs whole embryos (WE)-based genomic DNA preparation (the WE sequencing results are also provided here). Sequences show the successful C>T (G>A) conversion generating crispant tyr mutants that FS-derived genomic DNA can capture. The resulting dataset demonstrates the compatibility of the method developed with sequencing-based genotyping of early crispant embryos.   

Notes

To view the files common sequences analysis software are sufficient (Ape, Snapgene and Finch).

Funding provided by: Fondazione Umberto Veronesi
Crossref Funder Registry ID: http://dx.doi.org/10.13039/501100004710
Award Number: individual research fellowship

Funding provided by: H2020 Marie Skłodowska-Curie Actions
Crossref Funder Registry ID: http://dx.doi.org/10.13039/100010665
Award Number: 844636

Funding provided by: Agence Nationale de la Recherche
Crossref Funder Registry ID: http://dx.doi.org/10.13039/501100001665
Award Number: IHU FOReSIGHT ANR-18-IAHU-0001

Funding provided by: Fondation pour la Recherche Médicale*
Crossref Funder Registry ID:
Award Number: ECO20170637481

Funding provided by: Ligue Nationale Contre le Cancer*
Crossref Funder Registry ID:
Award Number: fellowship

Funding provided by: Ministero della Salute
Crossref Funder Registry ID: http://dx.doi.org/10.13039/501100003196
Award Number: Ricerca 5x1000

Funding provided by: Ministero della Salute
Crossref Funder Registry ID: http://dx.doi.org/10.13039/501100003196
Award Number: CCR-2017-23669081

Funding provided by: Ministero della Salute
Crossref Funder Registry ID: http://dx.doi.org/10.13039/501100003196
Award Number: RCR-2020-23670068_001

Funding provided by: Ministero della Salute
Crossref Funder Registry ID: http://dx.doi.org/10.13039/501100003196
Award Number: RCR-2021-23671215

Funding provided by: Ministero della Salute
Crossref Funder Registry ID: http://dx.doi.org/10.13039/501100003196
Award Number: GR-2019-12368907

Files

Raw_Sanger_Seq_Venditti_et_al.zip

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