2025-04-28 マサチューセッツ工科大学(MIT)
<関連情報>
- https://news.mit.edu/2025/gene-circuits-enable-more-precise-control-gene-therapy-0428
- https://www.cell.com/cell-systems/abstract/S2405-4712(25)00102-4
マイクロRNAに基づく遺伝子回路のモデル誘導設計により、多様な初代細胞への遺伝子導入カーゴの正確な投与が可能になる Model-guided design of microRNA-based gene circuits supports precise dosage of transgenic cargoes into diverse primary cells
Kasey S. Love ∙ Christopher P. Johnstone ∙ Emma L. Peterman ∙ Stephanie Gaglione ∙ Michael E. Birnbaum, ∙ Kate E. Galloway
Cell Systems Published:April 28, 2025
DOI:https://doi.org/10.1016/j.cels.2025.101269
Graphical abstract
Highlights
- ComMAND constrains the expression of output proteins across delivery methods
- ComMAND-regulated protein levels scale sublinearly with DNA dosage
- Modeling identifies physiological limits and tuning strategies for optimizing ComMAND
- ComMAND functions in primary cells and regulates clinically relevant genes
Summary
In a therapeutic context, supraphysiological expression of transgenes can compromise engineered phenotypes and lead to toxicity. To ensure a narrow range of transgene expression, we developed a single-transcript, microRNA-based incoherent feedforward loop called compact microRNA-mediated attenuator of noise and dosage (ComMAND). We experimentally tuned the ComMAND output profile, and we modeled the system to explore additional tuning strategies. By comparing ComMAND to two-gene implementations, we demonstrate the precise control afforded by the single-transcript architecture, particularly at low copy numbers. We show that ComMAND tightly regulates transgene expression from lentiviruses and precisely controls expression in primary human T cells, primary rat neurons, primary mouse embryonic fibroblasts, and human induced pluripotent stem cells. Finally, ComMAND effectively sets levels of the clinically relevant transgenes frataxin (FXN) and fragile X messenger ribonucleoprotein 1 (Fmr1) within a narrow window. Overall, ComMAND is a compact tool well suited to precisely specify the expression of therapeutic cargoes. A record of this paper’s transparent peer review process is included in the supplemental information.
