TY - JOUR
T1 - A Binary Cre Transgenic Approach Dissects Microglia and CNS Border-Associated Macrophages
AU - Kim, Jung-Seok
AU - Kolesnikov, Masha
AU - Peled-Hajaj, Shany
AU - Scheyltjens, Isabelle
AU - Xia, Yuan
AU - Trzebanski, Sebastien
AU - Haimon, Zhana
AU - Shemer, Anat
AU - Lubart, Alisa
AU - Van Hove, Hannah
AU - Chappell-Maor, Louise
AU - Boura-Halfon, Sigalit
AU - Movahedi, Kiavash
AU - Blinder, Pablo
AU - Jung, Steffen
N1 - We would like to thank all members of the Jung Laboratory, as well as the staff of the Weizmann Animal facility, the FACS facility, and the Weizmann Veterinary Services for expert technical help. We thank J. Hirrlinger and F. Kirchhoff for sharing split Cre reagents. A special thanks goes to S. Ben-Dor and R. Haffner for help with the CRISPR/Cas9 design and generation of the transgenic animals. S. Jung is the Incumbent of the Henry. H. Drake Professional Chair of Immunology. The Jung laboratory was supported by the Israel Science Foundation (887/11), the European Research Council (Adv ERC 340345), a collaborative network grant of the International Progressive MS Alliance (PMSA), the Minerva Foundation, the Deutsche Forschungsgemeinschaft (DFG) (CRC/TRR167 ‘NeuroMac’), the American Brain Foundation, the Roland N. Karlen Foundation, the Blythe Brenden-Mann Foundation, and the Estate of David Levinson. K. Movahedi was supported by Innoviris (Attract BB2B 2015-2), I. Scheyltjens and H. Van Hove are supported by FWO fellowships. Author Contributions J-S.K. and S.J. conceived the project and designed the experiments; J-S.K., Y.X., S.T., A.S., and S.B.-H. performed experiments; Z. H. helped with the bioinformatic analysis; L.C.-M. performed the bulk RNA-seq; M. K., A.L., S. P.-H., and P.B. performed imaging experiments; I. S, H.V.H., and K.M prepared the sc data set and performed bioinformatic analysis. J-S.K. and S.J. wrote the paper; S.J. supervised the project.
PY - 2021/1/12
Y1 - 2021/1/12
N2 - The developmental and molecular heterogeneity of tissue macrophages is unravelling, as are their diverse contributions to physiology and pathophysiology. Moreover, also given tissues harbor macrophages in discrete anatomic locations. Functional contributions of specific cell populations can in mice be dissected using Cre recombinase-mediated mutagenesis. However, single promoter-based Cre models show limited specificity for cell types. Focusing on macrophages in the brain, we establish here a binary transgenic system involving complementation-competent NCre and CCre fragments whose expression is driven by distinct promoters: Sall1ncre: Cx3cr1ccre mice specifically target parenchymal microglia and compound transgenic Lyve1ncre: Cx3cr1ccre animals target vasculature-associated macrophages, in the brain, as well as other tissues. We imaged the respective cell populations and retrieved their specific translatomes using the RiboTag in order to define them and analyze their differential responses to a challenge. Collectively, we establish the value of binary transgenesis to dissect tissue macrophage compartments and their functions.
AB - The developmental and molecular heterogeneity of tissue macrophages is unravelling, as are their diverse contributions to physiology and pathophysiology. Moreover, also given tissues harbor macrophages in discrete anatomic locations. Functional contributions of specific cell populations can in mice be dissected using Cre recombinase-mediated mutagenesis. However, single promoter-based Cre models show limited specificity for cell types. Focusing on macrophages in the brain, we establish here a binary transgenic system involving complementation-competent NCre and CCre fragments whose expression is driven by distinct promoters: Sall1ncre: Cx3cr1ccre mice specifically target parenchymal microglia and compound transgenic Lyve1ncre: Cx3cr1ccre animals target vasculature-associated macrophages, in the brain, as well as other tissues. We imaged the respective cell populations and retrieved their specific translatomes using the RiboTag in order to define them and analyze their differential responses to a challenge. Collectively, we establish the value of binary transgenesis to dissect tissue macrophage compartments and their functions.
UR - https://www.scopus.com/pages/publications/85098950772
U2 - 10.1016/j.immuni.2020.11.007
DO - 10.1016/j.immuni.2020.11.007
M3 - Article
C2 - 33333014
SN - 1074-7613
VL - 54
SP - 176
EP - 190
JO - Immunity
JF - Immunity
IS - 1
ER -