Open Dataset XSK1IXUI
Correlating the Structure and Gene Silencing Activity of Oligonucleotide-Loaded Lipid Nanoparticles Using Small-Angle X-Ray Scattering
Individual ASO-LNP samples and the screening sample plates were prepared by a robotic liquid handler-assisted, high-throughput solvent-injection method that we developed previously. Briefly, a liquid handler (TECAN EVO, NC, USA) was used to dispense the ASO dissolved in a citrate buffer (25 mM, pH 4) into a 96-well sample plate, as well as to mix individual lipid stocks at certain molar ratios to generate different lipid mixtures. For the PEG-lipid screening, LNPs were designed with the lipid composition with molar percent of MC3 : DSPC : cholesterol : PEG-lipids at 40 : 10 : (100 - X) : X, where X = 1, 3, or 5, a total lipid concentration of 1 mM, and a N/P ratio of 2. The prepared lipid mixtures were rapidly mixed with the ASO aqueous phase at a volume ratio of 1 : 3 (50 μL : 150 μL) in the sample plate using the robot, allowing for self-assembly of ASO-loaded LNPs, which were then characterized for particle structure by SAXS in the preparation buffer.In separate experiments, certain LNPs were processed through ultracentrifugation using an Amicon filter with the MWCO of 10 kD (Millipore, MA, USA) for purification and buffer exchange to PBS.Experimental description
SAXS data were collected in the high throughput mode (HT-SAXS) using the Advanced Light Source SIBYLS beamline 12.3.1 at the Lawrence Berkeley National Laboratory (CA, USA). X-ray wavelength was set at λ = 1.216 Å, and the sample-to-detector distance was 2070 mm, resulting in a scattering vector, q, ranging from 0.01 Å-1 to 0.45 Å-1. The scattering vector is defined as q = 4πsinθ/λ, where 2θ is the scattering angle. Experiments were performed at 20 °C as described elsewhere 29. Briefly, the sample was exposed for 10 s with the detector framing at 0.3 s to maximize the signal while merging the SAXS signal using the SAXS FrameSlice application (https://bl1231.als.lbl.gov/ran). No radiation damage was observed during the 10 s exposure, and all collected frames were merged.File description
PEGx_y.dat is a final merged SAXS profile , where x is a #ID of specific PEG-lipid and y is the molar ratio of PEG-lipid. \#1 DMPE (C14:0)-PEG0.55k - #2 DMPE(C14:0)-PEG1k #3 DMPE(C14:0)-PEG2k - #4 DPPE(C16:0)-PEG1k #5 DPPE(C16:0)-PEG2k - #6 DSPE(C18:0)-PEG0.55k #8 DSPE(C18:0)-PEG1k #9 DSPE(C18:0)-PEG2k - #10 DSPE(C18:0)-2arm-PEG2k- #11 DOPE(C18:1)-PEG0.55k #12 DOPE(C18:1)-PEG1k - #13 DOPE(C18:1)-PEG2k #14 DMG(C14:0)-PEG2k - #15 DSG(C18:0)-PEG2k #16 Ceramide(C8)-PEG0.75k #17 Ceramide(C8)-PEG2k - #18 Ceramide(C16)-PEG0.75k - #19 Ceramide(C16)-PEG2k
2023-02-08Data collection technique
1.216 ÅSample to Detector Distance
Lawrence Berkeley National Laboratory, The SIBYLS Beamline
United States of America
Data Download Terms:
Complete Set of SAS Data Files
The complete SAS dataset is downloadable as a zip file.
- LNP-ASO-PEGscreen.zip Download
Individual SAS Data Files (total 0)
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Supplemental Data and Supporting Materials (total 0)
These data and materials may include X-ray crystal structure coordinates, multi-angle light scattering data, .etc. It may also include additional details or methods pertaining to the SAXS experiments, or the researcher's interpretations of the results.
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