RNA Interference-Mediated Suppression of RAGE
- 1Institute of Biology of Aging and Medicine of Healthy Longevity with the Clinic of Predictive Medicine, Petrovsky National Research Centre of Surgery, 119435 Moscow, Russia
Abstract
The receptor for advanced glycation end products (RAGE/AGER) is an important mediator of inflammatory signaling and has been implicated in chronic low-grade inflammation associated with aging. Chronic low-grade inflammation (“inflammaging”) is a characteristic feature of biological aging, providing a rationale for investigating molecular regulators of inflammatory signaling such as RAGE. This study aimed to develop and validate short hairpin RNA (shRNA) constructs suppressing AGER/RAGE in human cell models. Five shRNA constructs targeting human AGER were designed, cloned into pLSL-puro, and verified by Sanger sequencing. Lentiviral vectors were produced in HEK293T cells and used to establish stable HEK293T, 786-O, and human umbilical vein endothelial cell (HUVEC) populations with AGER knockdown. AGER/RAGE expression was assessed by RT-qPCR, Western blotting, and flow cytometry. Silencing efficiency varied with the construct and the cellular model. sh1 produced the strongest suppression of AGER, exceeding 70% in selected models, whereas sh2 was effective in HEK293T and 786-O cells. Reduced RAGE expression was confirmed at the mRNA, protein, and cell-surface levels, identifying sh1 and sh2 as lead candidates. An AGER-focused workflow for lentiviral shRNA construct screening and experimental validation was established, providing a reproducible approach for generating stable cell models with reduced AGER/RAGE expression. Further work is required to confirm isoform specificity, assess off-target effects, and define the functional consequences of RAGE suppression.
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BibTeX
@article{muzaffarova2026interferencemediated,
title = {{RNA Interference-Mediated Suppression of RAGE}},
author = {Tatiana A. Muzaffarova and Elena M. Malinovskaya and Alexey Moskalev},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = oct,
doi = {10.3390/ijms27209167},
url = {https://www.mdpi.com/1422-0067/27/20/9167},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - RNA Interference-Mediated Suppression of RAGE
AU - Tatiana A. Muzaffarova
AU - Elena M. Malinovskaya
AU - Alexey Moskalev
PY - 2026
DA - 2026/10/16
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27209167
UR - https://www.mdpi.com/1422-0067/27/20/9167
AB - The receptor for advanced glycation end products (RAGE/AGER) is an important mediator of inflammatory signaling and has been implicated in chronic low-grade inflammation associated with aging. Chronic low-grade inflammation (“inflammaging”) is a characteristic feature of biological aging, providing a rationale for investigating molecular regulators of inflammatory signaling such as RAGE. This study aimed to develop and validate short hairpin RNA (shRNA) constructs suppressing AGER/RAGE in human cell models. Five shRNA constructs targeting human AGER were designed, cloned into pLSL-puro, and verified by Sanger sequencing. Lentiviral vectors were produced in HEK293T cells and used to establish stable HEK293T, 786-O, and human umbilical vein endothelial cell (HUVEC) populations with AGER knockdown. AGER/RAGE expression was assessed by RT-qPCR, Western blotting, and flow cytometry. Silencing efficiency varied with the construct and the cellular model. sh1 produced the strongest suppression of AGER, exceeding 70% in selected models, whereas sh2 was effective in HEK293T and 786-O cells. Reduced RAGE expression was confirmed at the mRNA, protein, and cell-surface levels, identifying sh1 and sh2 as lead candidates. An AGER-focused workflow for lentiviral shRNA construct screening and experimental validation was established, providing a reproducible approach for generating stable cell models with reduced AGER/RAGE expression. Further work is required to confirm isoform specificity, assess off-target effects, and define the functional consequences of RAGE suppression.
SN - 1422-0067
ER -