Sweet Dre-ams about intersectional genetics

Specific cell targeting with one site-specific recombinase is challenging. In this issue of Cell Stem Cell, Han et al., 2021Han X. Zhang Z. He L. Zhu H. Li Y. Pu W. M. Zhao Liu K. al.A suite new Dre drivers markedly expands the ability to perform intersectional genetic targeting.Cell Cell. 2021; 28: 1160-1176Abstract Full Text PDF PubMed Scopus (36) Google Scholar released a collection and demonstrate how two recombinases can be combined improve specificity lineage tracing gene inactivation in mice. Genetic conditional are approaches mouse genetics made possible by powerful Cre-lox (SSR) system (Sauer Henderson, 1988Sauer B. Henderson N. Site-specific DNA recombination mammalian cells Cre bacteriophage P1.Proc. Natl. Acad. Sci. USA. 1988; 85: 5166-5170Crossref (869) Scholar). Recently, second SSR has emerged that recognizes distinct target sites compares terms efficiency, namely Dre-rox (Anastassiadis 2009Anastassiadis Fu J. Patsch C. Hu S. Weidlich Duerschke Buchholz F. Edenhofer Stewart A.F. recombinase, like Cre, highly efficient E. coli, mice.Dis. Model. Mech. 2009; 2: 508-515Crossref (190) Hundreds lines have been generated express an under control specific achieve spatiotemporal over activity. Derivatives CreER DreER provide another level making activity dependent on administration tamoxifen. However, due complex dynamic nature vivo expression, it challenging find single driver truly type. As example involving tracing, there persistent questions regarding identity adipocyte progenitors generate adult fat (Guimarães-Camboa Evans, 2017Guimarães-Camboa Evans S.M. Are Perivascular Adipocyte Progenitors Mural Cells or Adventitial Fibroblasts?.Cell 2017; 20: 587-589Abstract (22) By logic once genetically encoded reporter activated label progenitor cell, all its cellular progeny must inherit label. When such reporters were Pdgfrb, was concluded some adipocytes arise from mural (vascular smooth muscle pericytes). But Pdgfrb although high many cells, not cells. Last year, shown labeled Tbx18-CreER Myh11-CreER extremely inefficient at generating variety pro-adipogenic conditions. Instead, fibroblastic Pdgfra-CreER principal source (Cattaneo 2020Cattaneo P. Mukherjee D. Spinozzi Larcher V. Stallcup W.B. Kataoka Chen Dimmeler al.Parallel Lineage-Tracing Studies Establish Fibroblasts as Prevailing Vivo Progenitor.Cell reports. 2020; 30: 571-582.e2Abstract (30) Notably, study (Sun 2020Sun Sakashita Kim Tang Upchurch G.M. Yao Berry W.L. Griffin T.M. Olson L.E. Mosaic Mutant Analysis Identifies PDGFRalpha/PDGFRbeta Negative Regulators Adipogenesis.Cell 26: 707-721.e5Abstract (20) Scholar) Flp sequence mosaic analysis Pdgfra mutant progenitors, which confirmed Pdgfra-expressing identified function PDGF receptors inhibitors adipogenesis. current al. (Han revisited question using double-recombinase approach Dre. While previous studies compared different mice, here, they performed dual-reporter same mouse. For this, created Pdgfra-DreER red fluorescent mice crossed them Pdgfrb-CreER green They used tamoxifen-inducible screen organs for three populations cells: Pdgfra+ (red), Pdgfrb+ (green), Pdgfra+Pdgfrb+ (yellow). Shortly after activating tamoxifen, each organ contained proportion colors, depicted yellow adventitial fibroblasts localized around vascular structures. Then, exposing three-color challenges induce formation found but marked respective color. This result parallels conclusions Cattaneo Sun Scholar, further reinforcing notion fibroblast-like primary tissues. If elegant brings mind other about lineages overlap vivo, resources offered dream come true. Besides Pdgfra-DreER, 71 knockin expressing 56 genes. these tools might applied questions, Cre-based skeletal stem (SSCs) heterogeneous respect potential (Serowoky 2020Serowoky M.A. Arata C.E. Crump J.G. Mariani F.V. Skeletal insights into maintaining regenerating skeleton.Development. 147: dev179325Crossref Several controlled genes mark putative SSCs, including Axin2-2A-DreER, Gli1-DreER, Lepr-2A-DreER, Sox9-2A-DreER. Any SSC could existing compare adipocyte, osteoblast, chondrocyte, stromal potentials skeleton. also demonstrated ways sequentially increase tracing. recognized selectively subsets adipocytes, widely storing white (Adipoq-CreER relatives) targets thermogenic brown adipocytes. To more specifically tools: Ucp1 (Ucp1-Dre), Dre-inactivating pan-adipocyte Plin1 (Plin1-rox-CreER-rox). together following NOT-logic strategy, Plin1-driven suppressed Another problem currently exists lymphatic endothelial (LECs) because LECs (Prox1-CreER neurons, hepatocytes, cardiomyocytes, lens LECs, pan-endothelial Cdh5 (Cdh5-Dre) combination Dre-dependent Prox1 (Prox1-rox-STOP-rox-CreER). Consequently, Prox1-driven expressed AND-logic strategy. future, finally solve century-old controversy non-venous origins (Martinez-Corral 2015Martinez-Corral I. Ulvmar M.H. Stanczuk Tatin Kizhatil John S.W. Alitalo Ortega Makinen T. Nonvenous origin dermal vasculature.Circ. Res. 2015; 116: 1649-1654Crossref (156) Scholar; Maruyama 2019Maruyama Miyagawa-Tomita Mizukami Matsuzaki Kurihara Isl1-expressing contributes cardiac vessel development.Dev. Biol. 2019; 452: 134-143Crossref (37) Srinivasan 2007Srinivasan R.S. Dillard M.E. Lagutin O.V. Lin F.J. Tsai M.J. Samokhvalov I.M. Oliver G. Lineage demonstrates venous vasculature.Genes Dev. 2007; 21: 2422-2432Crossref (387) Lastly, proof-of-concept experiments inactivate Plin1+ Prox1+ LECs. Plin1-NOT-Ucp1 deleted Pparg, Pten, Ctnnb1 Using Prox1-AND-Cdh5 RNA-modifying enzyme Mettl3, embryonic cases, RNA and/or protein levels floxed depleted increased tissue when Dre/Cre knockouts Cre-only knockouts. There now large drivers, yet much work lies ahead realize full Dre-rox. Eight 70+ built lox-STOP-lox cassette make Cre-dependent (LSL-Dre). These useful Cre-AND-Dre lack rox-flanked means LSL-Dre cannot deletion time. The lines, Plin1-rox-CreER-rox Prox1-rox-STOP-rox-CreER, available lox-flanked greater specificity, experiments. only collection. Therefore, widespread use will depend individual researchers, consortium, construct alleles rox-CreER-rox, rox-STOP-rox-CreER, cleverly designed cassettes. Nevertheless, valuable may beginning true those trying answer historically difficult through genetics. A targetingHan al.Cell CellFebruary 9, 2021In BriefThe combinatory recombinase-mediated significantly enhances precision targeting. developed than 70 diverse lineages. Highlighting their application, perivascular knockout (WAs) (LECs). Full-Text Open Archive