Fig. 1: Development of DIRECTED.

a Schematic of lentiviral vector production and lentivector-mediated delivery to target cells. b Schematic to expand tropism by co-expressing VSV-G and a variant of protein AG (pAG) that can recruit an antibody on virions (left). Transduction efficiency of HEK293FT cells (as a percentage of mCherry+ cells) of lentiviral particles with various ratios of VSV-G and pAG in the presence or absence of αHLA-A2 (right, N = 4 for all conditions, p-values: 0.03042, 0.00162, 0.001386, 0.0001269, 7.39E-05, 0.001728, 0.01287, 1, 1). c Schematic to reduce VSV-G-mediated tropism by mutating residues that interact with low-density lipoprotein receptors (LDL-R) (left). Quantification of titers for VSV-G mutants (middle; N = 18 for WT; 12 for H8A, K47Q, Y209A, R354Q; 6 for H8A&K47Q, H8A&R354Q, K47Q&R354Q, R354Q&Y209A; p-values: 1, 2.25E-07, 3.22E-06, 1.92E-07, 1.36E-06, 5.70E-05, 7.31E-07, 6.03E-07). Transduction efficiency of HEK293FT cells (as a percentage of mCherry+ cells) of lentiviral particles with VSV-G point mutations (right; N = 6 for WT; 4 for H8A, K47Q, Y209A, R354Q; 2 for H8A&K47Q, H8A&R354Q, K47Q&R354Q, R354Q&Y209A; p-values: 2.28E-07, 4.74E-08, 0.016, 5.26E-07, 5.16E-07, 5.22E-07, 5.15E-07, 4.98E-07). d Schematic showing separate fusion and targeting components by combining VSV-G K47Q/R354Q (VSV-Gdm) with pAG (left). Transduction efficiency of HEK293FT wild-type cells or B2M knockout cells as a percentage of mCherry+ cells (right, N = 4 for all conditions) of lentiviral particles with VSV-Gdm and pAG in the presence or absence of αHLA-A2 at different MOI. (p-values: wild-type: 0.0001926, 1.15E-06, 1.54E-06, 0.002304, 1.19E-05, 0.0001458; ΔB2M: 1, 0.258, 0.876, 0.21, 0.822, 0.02598). For panels b, c, and d, a two-sided Welch’s t-test with Bonferroni correction was used. [ns, not significant; * p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; Data are presented as mean ± standard deviation. Source data are provided as a Source Data file].