title: Engineered red blood cells as carriers for systemic delivery of a wide array of functional probes.
abstract: We developed modified RBCs to serve as carriers for systemic delivery of a wide array of payloads. These RBCs contain modified proteins on their plasma membrane, which can be labeled in a sortase-catalyzed reaction under native conditions without inflicting damage to the target membrane or cell. Sortase accommodates a wide range of natural and synthetic payloads that allow modification of RBCs with substituents that cannot be encoded genetically. As proof of principle, we demonstrate site-specific conjugation of biotin to in vitro-differentiated mouse erythroblasts as well as to mature mouse RBCs. Thus modified, RBCs remain in the bloodstream for up to 28 d. A single domain antibody attached enzymatically to RBCs enables them to bind specifically to target cells that express the antibody target. We extend these experiments to human RBCs and demonstrate efficient sortase-mediated labeling of in vitro-differentiated human reticulocytes.
authors: Whitehead Institute for Biomedical Research, Cambridge, MA 02142; and.
authors: Jiahai
authors: J
authors: Shi
authors: Whitehead Institute for Biomedical Research, Cambridge, MA 02142; and.
authors: Lenka
authors: L
authors: Kundrat
authors: Departments of Biological Engineering and.
authors: Novalia
authors: N
authors: Pishesha
authors: Whitehead Institute for Biomedical Research, Cambridge, MA 02142; and.
authors: Angelina
authors: A
authors: Bilate
authors: Whitehead Institute for Biomedical Research, Cambridge, MA 02142; and.
authors: Chris
authors: C
authors: Theile
authors: Whitehead Institute for Biomedical Research, Cambridge, MA 02142; and.
authors: Takeshi
authors: T
authors: Maruyama
authors: Whitehead Institute for Biomedical Research, Cambridge, MA 02142; and.
authors: Stephanie K
authors: SK
authors: Dougan
authors: Whitehead Institute for Biomedical Research, Cambridge, MA 02142; andBiology, Massachusetts Institute of Technology, Cambridge, MA 02139 ploegh@wi.mit.edu lodish@wi.mit.edu.
authors: Hidde L
authors: HL
authors: Ploegh
authors: Whitehead Institute for Biomedical Research, Cambridge, MA 02142; andDepartments of Biological Engineering andBiology, Massachusetts Institute of Technology, Cambridge, MA 02139 ploegh@wi.mit.edu lodish@wi.mit.edu.
authors: Harvey F
authors: HF
authors: Lodish
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D060846:Cell Engineering; D002478:Cells, Cultured; D004900:Erythroblasts / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D012156:Reticulocytes / Q000378:metabolism*
keywords: mammalian cell engineering; sortaggable GPA and Kell; survival time in circulation