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. 2019 Oct 23;47(21):11441–11451. doi: 10.1093/nar/gkz914

Figure 2.

Figure 2.

Preventing aggregation by adjacent ssDNA overhangs. (A) PAGE showing cholesterol-modified ssDNA (ss chol, blue) on its own and hybridized to a complementary strand (orange) of varying overhang length (–6 to +10 nt with respect to the chol-DNA strand). L denotes a DNA ladder. (B–E) Instantaneous snapshots of all-atom MD simulations of the DNA duplex with one cholesterol tag with either no overhang (B and C) or a 6 nt overhang (D and E), each shown at the beginning (0 μs) and end (0.5 μs) of the equilibrium MD simulation. Close-ups illustrate that cholesterol is more freely accessible without the overhang (C) but shielded by the ssDNA if the overhang is present (E) (Supplementary Movies M2 and M3). (F) Cy3-labeled duplex with +6 nt overhang (red) shows attachment to droplet-stabilized giant unilamellar vesicles (brightfield) (61). The cholesterol-shielding overhang does not inhibit lipid membrane interaction. Scale bars denote 50 μm. For images of Atto488-labeled lipids see Supplementary Figure S11.