1. Fig. 1 Strategy for construction of T4-AAV hybrid vectors and schematic for delivery into mammalian cells.
Strategy for construction of T4-AAV hybrid vectors and schematic for delivery into mammalian cells. (A) T4 DNA packaging and protein display machine. The zoomed oval view below indicates the DNA packaging motor gp17 (pentamer) and portal gp20 (dodecamer). Enlarged capsomer (hexamer) shows the arrangement of the major capsid protein gp23* (cyan), Soc trimers (blue), and Hoc fiber (yellow). (B) Sixty AAV DJ subunits assemble to form the icosahedrally symmetric virus capsid [Protein Data Bank (PDB) 3J1Q]. The T4 (120 nm × 86 nm) and AAV (25 nm) capsids are shown approximately to scale. (C and D) Design principles of the T4-AAV vector. Soc-biotin-avidin (SBA) (C) and Hoc–biotin acceptor peptide (BAP)–biotin–avidin (HBBA) (D) are applied to conjugate the T4 head with biotin-labeled AAV via the avidin-biotin interaction. The protein ribbon structures and biotin structure in the rectangle depict the detailed SBA and HBBA linkers. The ribbon diagrams at the bottom show Hoc and Soc structures with exposed N and C termini (PDB 3SHS and PDB 3IG9). (E) Schematic representation of protein and gene delivery into mammalian cells by the T4-AAV vector. I, binding to receptors on the cell membrane; II, endocytosis; III, endosomal/lysosomal escape; IV, uncoating to release protein cargo; V, T4-encapsidated DNA release; VI, transport of DNAs into the nucleus along microtubules; VII, transcription of delivered genes; VIII and IX, mRNA transport into cytosol and protein production. ssDNA, single-stranded DNA; dsDNA, double-stranded DNA; ITRs, inverted terminal repeats.
Jingen Zhu et al. Sci Adv 2019;5:eaax0064
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