Genetic Tools to Study Cardiovascular Biology

Abstract

Progress in biomedical science is tightly associated with the improvement of methods and genetic tools to manipulate and analyze gene function in mice, the most widely used model organism in biomedical research. The joint effort of numerous individual laboratories and consortiums has contributed to the creation of a large genetic resource that enables scientists to image cells, probe signaling pathways activities, or modify a gene function in any desired cell type or time point, à la carte. However, as these tools significantly increase in number and become more sophisticated, it is more difficult to keep track of each tool's possibilities and understand their advantages and disadvantages. Knowing the best currently available genetic technology to answer a particular biological question is key to reach a higher standard in biomedical research. In this review, we list and discuss the main advantages and disadvantages of available mammalian genetic technology to analyze cardiovascular cell biology at higher cellular and molecular resolution. We start with the most simple and classical genetic approaches and end with the most advanced technology available to fluorescently label cells, conditionally target their genes, image their clonal expansion, and decode their lineages.

Multicolor strategies for investigating clonal expansion and tissue plasticity

“…Pontes-Quero and colleagues developed an elegant genetic strategy for the conditional induction of mosaic gene expression based on the expression of distinct and compatible FP: inducible, fluorescent, and genetic mosaic (ifgMosaic) [ 33 ], reviewed in [ 34 ]. This system, which can lead up to 15 distinct clone colors, relies on the combination of Cre/lox recombination and knock-in insertions in the mouse Rosa26 locus of multiFP transgenes, is compatible with live imaging and immunostaining, and has the possibility to insert a mosaic co-expression of other genes of interest as well (Fig.…”

A genome-wide library of MADM mice for single-cell genetic mosaic analysis

“…With the availability of MADM for all mouse autosomes, the phenomenon of cell competition can be studied holistically and for virtually any candidate gene function associated with it in diverse biological contexts in health and disease. Although MADM technology is versatile and offers multiple applications (discussed above), the method also has certain limitations and alternative systems may be considered depending on the biological question and context (Garcia- Gonzalez et al, 2020;Hansen and Hippenmeyer 2020). For instance, the expression of the two markers relies on a single-copy insertion and thus the expression level may be lower than, for example, that in viralbased systems.…”

Partial endothelial-to-mesenchymal transition (EndMT) contributes to lumen re-organization after carotid artery ligation

“…Technological advances in genetically engineered mice enable us to analyze the fate of vascular cells during vascular injury 28 . Tracing of the endothelial lineage using the constitutively active systems such as Tie2Cre mice has shown Cre-mediated recombination in endothelial-derived cells and circulating leukocytes 11 .…”