Journal:

Article Title: Models of primitive cellular life: polymerases and templates in liposomes

doi: 10.1098/rstb.2007.2066

Figure Lengend Snippet: Accumulation of RNA within liposomes over time. RNA polymerization within 400 nm (filled diamonds) and 800 nm (open squares) extruded liposomes was assayed by fluorescence (RiboGreen assay) at various times during 70 temperature cycles.

Article Snippet: RiboGreen RNA fluorescent staining was used to determine the RNA product of the entrapped reaction, and the fluorescence was measured with an SLM 8000C spectrofluorimeter (SLM Instruments, Inc.).

Techniques: Fluorescence

Journal:

Article Title: Models of primitive cellular life: polymerases and templates in liposomes

doi: 10.1098/rstb.2007.2066

Figure Lengend Snippet: DNA-template encapsulation yields and amount of RNA (RiboGreen assay) produced within DMPC liposomes after 70 temperature cycles as function pore size of the filters used for the extrusion

Article Snippet: RiboGreen RNA fluorescent staining was used to determine the RNA product of the entrapped reaction, and the fluorescence was measured with an SLM 8000C spectrofluorimeter (SLM Instruments, Inc.).

Techniques: Produced

Journal: bioRxiv

Article Title: Modular mRNA lipid nanoparticle platform rescues diverse genetic male infertility

doi: 10.1101/2025.11.13.688269

Figure Lengend Snippet: (a) Upper: Schematic of the Ai9 Cre-reporter system. In Ai9 mice, a LoxP-flanked STOP cassette prevents tdTomato expression until Cre recombinase excises the STOP, activating red fluorescence in any cell that has received Cre. Lower: In vivo fluorescence imaging of Ai9 mice 48 h after intratubular injection of Cre mRNA LNP (n = 3 mice per group). H, heart; Lu, lung; Li, liver; S, spleen; K, kidney; T, testis. (b) A seminiferous tubule cross-section from a treated testis, co-stained for tdTomato (red), the Sertoli cell marker SOX9 (green), and nuclei (DAPI, blue). Scale bar: 40 µm. (c) Upper: Gating strategy to resolve stages: pre-leptotene spermatogonia (2N, within P5), leptotene/zygotene spermatocytes (replicating DNA, P6), pachytene spermatocytes (4N, mid-meiosis, P7), diplotene spermatocytes (4N, late meiosis, P8), secondary spermatocytes (2N, post-meiosis II, P9) and round spermatids (1N, P10). Lower: Percentage of tdTomato-positive cells within each gated population for ALC-0315, MC3 versus SM-102 LNP–Cre-treated testes (48 h post-injection; n = 6 testes each). (d) Kinetics of eGFP expression in testes following delivery of different RNA formats. Mice were injected (via MC3 LNP) with either N¹mΨ-modified eGFP mRNA (red), self-amplifying RNA replicon (saRNA, black) or circular RNA (circRNA, blue) encoding eGFP. Plotted is the radiant efficiency (GFP fluorescence) of the injected testis over 14 days (mean ± s.d.; n = 3 mice per group). (e) Time-course comparison of unmodified versus pseudouridine-modified mRNA (both encoding eGFP). Unmodified mRNA (black) drives an early peak of GFP (within 1 day) followed by a steep decline; modified mRNA (red) shows a delayed peak (∼3 days) and a slower decline (mean ± s.d.; n = 3 mice per group). (f) RT-qPCR measurement of eGFP mRNA in the testis at various times after injection of unmodified versus modified eGFP mRNA. (g) Innate immune gene induction in the testis by unmodified versus modified mRNA. RT-qPCR measurements were taken at 6, 12 and 24 h after injection of 10 µg eGFP mRNA (either unmodified or N¹mΨ-modified). Data are mean ± s.d. (n = 3 mice per group).

Article Snippet: Encapsulation efficiency of mRNA was assessed with a RiboGreen RNA fluorescence assay (Thermo Fisher Quant-iTTM RiboGreen kit), by treating aliquots of the LNP with or without 0.2% Triton X-100 to distinguish encapsulated vs. total RNA.

Techniques: Expressing, Fluorescence, In Vivo, Imaging, Injection, Staining, Marker, Modification, Comparison, Quantitative RT-PCR