iFlpMosaics enable the multispectral barcoding and high-throughput comparative analysis of mutant and wild-type cells
To understand gene function, it is necessary to compare cells carrying the mutated target gene with normal cells. In most biomedical studies, the cells being compared are in different mutant and control animals and, therefore, do not experience the same epigenetic changes and tissue microenvironment. The experimental induction of genetic mosaics is essential to determine a gene cell-autonomous function and to model the etiology of diseases caused by somatic mutations. Current technologies used to induce genetic mosaics in mice lack either accuracy, throughput or barcoding diversity. Here we present the iFlpMosaics toolkit comprising a large set of new genetic tools and mouse lines that enable recombinase-dependent ratiometric induction and single-cell clonal tracking of multiple fluorescently labeled wild-type and Cre-mutant cells within the same time window and tissue microenvironment. The labeled cells can be profiled by multispectral imaging or by fluorescence-activated flow cytometry and single-cell RNA sequencing. iFlpMosaics facilitate the induction and analysis of genetic mosaics in any quiescent or progenitor cell, and for any given single or combination of floxed genes, thus enabling a more accurate understanding of how induced genetic mutations affect the biology of single cells during tissue development, homeostasis and disease.
Authors
- Federica Francesca Lunella (ORCID: https://orcid.org/0000-0002-8943-6032)
- Carlos Torroja (ORCID: https://orcid.org/0000-0001-8914-3400)
- Maria S. Sanchez-Muñoz (ORCID: https://orcid.org/0000-0002-7661-2295)
- Irene García-Gónzalez (ORCID: https://orcid.org/0000-0001-8508-8344)
- Ivana Zagorac
- Stefano Gambera (ORCID: https://orcid.org/0000-0003-2998-8502)
- Lourdes García-Ortega (ORCID: https://orcid.org/0000-0002-9696-5047)
- Wen Luo (ORCID: https://orcid.org/0000-0002-5856-3733)
- Rui Benedito (ORCID: https://orcid.org/0000-0003-2458-3055)
- Macarena Fernández-Chacón (ORCID: https://orcid.org/0000-0001-5831-0923)
- Verónica Casquero-García (ORCID: https://orcid.org/0000-0002-2365-394X)
- Álvaro Regano (ORCID: https://orcid.org/0000-0002-2143-3011)
- Luis Diago-Domingo
- Mariya Lytvyn
- Macarena De Andrés-Laguillo
- Aroa Garcia-Cabero (ORCID: https://orcid.org/0000-0003-4008-6243)
- Fátima Sánchez-Cabo
- Silvia Faria da Rocha (ORCID: https://orcid.org/0000-0002-8001-1898)
Institutions
- Spanish National Centre for Cardiovascular Research (ES)
- Spanish National Centre for Cardiovascular Research (ES)
Publication Details
- Journal
- Nature Methods
- Published
- 2024-12-13
- DOI
- https://doi.org/10.1038/s41592-024-02534-w
- Citations
- 4
- Primary Topic
- Cell Image Analysis Techniques
- Type
- article
- Field-Weighted Citation Impact
- 2.65
Funders
- Ministerio de Ciencia, Innovación y Universidades
- Ministerio de Economía y Competitividad
- Centro Nacional de Investigaciones Cardiovasculares
- University of Texas MD Anderson Cancer Center
- Instituto de Salud Carlos III
- Agencia Estatal de Investigación