A team at the Centro Nacional de Investigaciones Cardiovasculares (CNIC) has developed a comprehensive set of innovative genetic tools and mouse lines, called iFlpMosaics, designed to enhance the study of gene function and its implications in health and disease.
The groundbreaking study, led by Dr. Rui Benedito and published in Nature Methods, presents a pioneering approach that overcomes critical limitations of existing methods for generating genetic mosaics. These innovations will enable scientists to more accurately investigate the effects of somatic mutations on cellular biology and disease.
The study highlights the iFlpMosaics toolkit's utility across different experimental setups, detailing how it allows scientists to track the effects of single or multiple gene deletions within the same tissue. This advance opens the way to deeper insight into the function of genes in cell biology, regeneration, and disease.
Understanding gene function is pivotal for the progress of biomedical research. Traditional biomedical genetic studies compare cells from distinct mutant and control animals, a method that often fails to account for the differing epigenetic landscapes and tissue microenvironments within each animal. "This disparity can lead to confusing results, complicating the interpretation of gene function," explained Dr. Benedito.
The iFlpMosaics toolkit is unburdened by these shortcomings and allows researchers to induce genetic mosaics with high throughput and precision, making it easier to study cell-autonomous gene function directly within the same organism.
"Our work with these new genetic tools highlights the importance of generating genetic mosaics from identical progenitor cells, within the same animal, if we want to fully understand the function of different genes in multiple cell types during organ development or in disease models" said Dr. Irene García González, the first author on the study.
Current technologies for inducing genetic mosaics, such as MADM (Mosaic Analysis with Double Markers) or Cre-dependent mosaics, are hampered by technical issues related to low efficiency or reliability. The iFlpMosaics toolkit overcomes these issues, offering a robust platform for the ratiometric induction and clonal tracking of fluorescently labeled wildtype and mutant cells.
The toolkit not only enhances the understanding of genetic mutations in tissue development and disease processes, but also facilitates the study of complex interactions between cells within their microenvironment.
"iFlpMosaics offers a big step forward for researchers studying diseases caused by somatic mutations, such as cancer and vascular malformations" said Dr. Rui Benedito. "It's precision and versatility provide an important resource for anyone seeking a better understanding of gene function in normal organ development and function, as well as in disease settings."
The study was funded by the European Research Council (ERC) through Starting Grant AngioGenesHD (638028) and Consolidator Grant AngioUnrestUHD (101001814), the Spanish Ministry of Science, Innovation, and Universities (SAF2017-89299-P y PID2020-120252RB-I00), and the la Caixa Foundation (HR19-00120 and HR22-00316 AngioHeart).
About the CNIC
The CNIC is an affiliate center of the Carlos III Health Institute (ISCIII), an executive agency of the Spanish Ministry of Science, Innovation, and Universities. Directed by Dr. Valentín Fuster, the CNIC is dedicated to cardiovascular research and the translation of the knowledge gained into real benefits for patients. The CNIC has been recognized by the Spanish government as a Severo Ochoa center of excellence (award CEX2020-001041-S, funded by MICIN/AEI/10.13039/501100011033). The center is financed through a pioneering public-private partnership between the government (through the ISCIII) and the Pro-CNIC Foundation, which brings together 11 of the most important Spanish private companies.
