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FTT5
Formulation & Performance
Formulation & Performance
Study-specific compositions, preparation conditions and experimental outcomes for FTT5.
Full article and complete SI reviewed; liver reporter, FVIII replacement, base editing and lipid clearance are separated.
1. Formulations and component amounts
Reported composition and calculated mass are separate. Molecular-weight inputs use the current catalog values; DMG-PEG2000 is fixed initially at 2507. Edit inputs only when using a verified alternative material.
Original in vivo FTT5 recipe
molar parts (sum 90.75)Main formulation Methods
| Component | Molecular weight | Molar parts | Normalized mol% | Amount (µmol) | Weight (mg) |
|---|---|---|---|---|---|
| FTT5 | 22.039 | 25.1735 | 50.0720 | ||
| DOPE | 33.058 | 37.7602 | 28.0947 | ||
| Cholesterol | 44.077 | 50.3469 | 19.4666 | ||
| DMG-PEG2000 | 0.826 | 0.9440 | 2.3666 |
Calculated total: 100.0000 mg lipid. RNA, solvent and process loss are excluded.
The mass worksheet uses the existing catalog molecular-weight input (1989.07791852951).
How component weights are calculated
For total lipid mass M (mg), molar parts rᵢ and molecular weight MWᵢ: component mass = M × rᵢ × MWᵢ / Σ(r × MW). Component amount (µmol) = component mass (mg) × 1000 / MWᵢ. Molar parts are normalized only for the displayed percentage. The calculation does not infer N/P or RNA dose.
2. Preparation and experimental setup
| Parameter | Reported procedure / limitation | Source |
|---|---|---|
| Assembly | Lipids dissolved in ethanol; RNA in citrate buffer. Pipette mixing for in vitro tests; microfluidic formulation for in vivo experiments and cryo-TEM. | Materials and Methods |
| Unspecified process values | The reviewed protocol does not establish an exact citrate pH, flow ratio, total flow or N/P for this recipe. | Not filled from another lipid protocol |
| Optimization outcome | Some optimized recipes performed better in vitro but worse in vivo. The original composition was retained for subsequent in vivo work. | SI Figs. S4–S5 |
| Cargo mapping | FVIII mRNA and adenine-base-editor mRNA/sgRNA were studied in separate dose and endpoint experiments. | Main Figs. 3–4 |
3. Particle characterization
| Measurement / batch | Reported result | Evidence context |
|---|---|---|
| Original FTT5 LNP | Approximately 100 nm; PDI <0.2; zeta potential approximately −11 mV; encapsulation approximately 91% | Main Fig. 2 |
| Molar-ratio basis | The reported entries sum to 90.75 parts | Normalized only for the mass calculation |
| Neat-lipid shelf life | Not established by formulated-particle or clearance experiments | Follow the supplied batch documentation |
4. Performance by experiment
| Experiment and dose | Readout and comparator | Result and source |
|---|---|---|
| Hepatic reporter expression C57BL/6 mice; intravenous Fluc mRNA 0.5 mg/kg | Readout: 6 h Comparator: TT3 formulation | Approximately twofold greater liver expression; the reported organ signal is normalized by tissue weight.Main Fig. 2; n=2 |
| Lipid clearance Intravenous RNA dose 1.8 mg/kg; lipid amounts measured in liver | Readout: 20 min, 6 h, 24 h and 48 h Comparator: FTT9 and other study formulations | FTT5 retained approximately half of its liver lipid amount at 48 h. The >80% removal at 6 h and complete removal by 24 h belong to FTT9, not FTT5.Main Fig. 2; n=3 |
| FVIII protein supplementation Wild-type and hemophilia-A mice; FVIII mRNA 2 mg/kg IV | Readout: 6 h and 12 h Comparator: Study control groups | FVIII concentrations exceeded 140 ng/mL in wild-type mice and 220 ng/mL in hemophilia-A mice at the reported points. These are protein-expression endpoints.Main Fig. 3; n=3 |
| FVIII functional activity Hemophilia-A mice; same 2 mg/kg FVIII-mRNA study | Readout: 6 h and 12 h Comparator: 1 IU/mL defined as 100% activity standard | Reported activity approximately 73% and 90%, respectively; not a percentage of corrected liver cells.Main Fig. 3; n=3 |
| Adenine base editing BALB/c mice; ABE mRNA:sgRNA 1:1 by mass; intravenous total-RNA dose response | Readout: Day 7 targeted sequencing Comparator: TT3 comparison and dose-response controls | Editing plateaued near 60% at 1 mg/kg. The reported ED50 <0.125 mg/kg describes the dose-response fit, not necessarily 50% absolute editing.Main Fig. 4; n=5; SI Fig. S7 |
| Short-term histology Hemophilia-A mice; FVIII mRNA 2 mg/kg IV | Readout: 72 h Comparator: Study controls | No evident abnormalities were reported in examined sections. Repeat-dose, long-term and human tolerability are not established by this endpoint.Main Fig. 3 / SI safety assessment |
Mechanistic evidence
Inhibitor experiments in Hep3B cells reported reduced uptake with several pathway inhibitors; calcein-release observations support endosomal escape. These assays do not establish an exclusive entry mechanism.
5. Interpretation and unresolved details
- Do not assign FTT9 rapid-clearance values to FTT5.
- Improved cell-culture activity after reformulation did not translate into improved in vivo expression.
- Adenine editing, FVIII protein concentration and coagulation activity use different denominators and cannot be combined as one efficacy percentage.
Results apply to the stated LNP composition, cargo, process, dose, route and model. LNP experimental measurements are not batch-release specifications for the isolated lipid.
6. References and data access
Zhang X, Zhao W, Nguyen GN, et al. Functionalized lipid-like nanoparticles for in vivo mRNA delivery and base editing. Science Advances. 2020;6(34):eabc2315.
Download experiment table (CSV) · Open printable Formulation & Performance guide
Revision FP-20261022 · Source review: 22 October 2026 · DC Chemicals