Construction and identification of RNA interference lentiviral vector for rat disruptor of telomeric silencing 1-like gene

doi:10.12307/2026.825

Chinese Journal of Tissue Engineering Research ›› 2026, Vol. 30 ›› Issue (31): 8197-8204.doi: 10.12307/2026.825

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Construction and identification of RNA interference lentiviral vector for rat disruptor of telomeric silencing 1-like gene

Yang Zishu1, 2, 3, Yang Jian2, 3, 4, 5, Huang Cuiyuan2, 3, 4, Cheng Chen2, 3, 4, Wu Di2, 3, 4, Zhang Jing2, 3, 4, 5

1Department of Ultrasound Imaging, 4Central Laboratory, 5Department of Cardiology, The First College of Clinical Medical Science, China Three Gorges University & Yichang Central People's Hospital, Yichang 443003, Hubei Province, China; 2Hubei Key Laboratory of Ischemic Cardiovascular Disease, Yichang 443003, Hubei Province, China; 3Hubei Provincial Clinical Research Center for Ischemic Cardiovascular Disease, Yichang 443003, Hubei Province, China

Received:2025-09-26 Accepted:2026-01-06 Online:2026-11-08 Published:2026-05-25
Contact: Zhang Jing, MD, Associate researcher, Hubei Key Laboratory of Ischemic Cardiovascular Disease, Yichang 443003, Hubei Province, China; Hubei Provincial Clinical Research Center for Ischemic Cardiovascular Disease, Yichang 443003, Hubei Province, China; Central Laboratory, and Department of Cardiology, The First College of Clinical Medical Science, China Three Gorges University & Yichang Central People's Hospital, Yichang 443003, Hubei Province, China
About author:ang Zishu, MS, Attending physician, Department of Ultrasound Imaging, The First College of Clinical Medical Science, China Three Gorges University & Yichang Central People's Hospital, Yichang 443003, Hubei Province, China; Hubei Key Laboratory of Ischemic Cardiovascular Disease, Yichang 443003, Hubei Province, China; Hubei Provincial Clinical Research Center for Ischemic Cardiovascular Disease, Yichang 443003, Hubei Province, China Yang Jian, MD, Professor, Chief physician, Hubei Key Laboratory of Ischemic Cardiovascular Disease, Yichang 443003, Hubei Province, China; Hubei Provincial Clinical Research Center for Ischemic Cardiovascular Disease, Yichang 443003, Hubei Province, China; Central Laboratory, and Department of Cardiology, The First College of Clinical Medical Science, China Three Gorges University & Yichang Central People's Hospital, Yichang 443003, Hubei Province, China Yang Zishu and Yang Jian contributed equally to this article.
Supported by:
National Natural Science Foundation of China, No. 82170418 (to ZJ); National Natural Science Foundation of China, No. 82271618, 82471616 (to YJ); Hubei Provincial Natural Science Foundation Innovation Group Project, No. 2022CFA015 (to YJ); Hubei Provincial Key Research and Development Program, No. 2023BCB139 (to YJ); Hubei Provincial Regional Science and Technology Innovation Program, No. 2025EIA015 (to ZJ); Hubei Provincial Science and Technology Innovation Platform Project, No. 2025CCB016 (to HCY)

Abstract

Abstract: BACKGROUND: Disruptor of telomeric silencing 1-like (DOT1L) is a key methyltransferase that catalyzes histone H3 lysine 79 methylation, and its aberrant function is associated with vascular remodeling and a variety of diseases. The construction of stable DOT1L ribonucleic acid interference (RNAi) lentiviral vectors can provide an effective experimental tool to study the biological function of DOT1L in cardiovascular diseases and related disease mechanisms.
OBJECTIVE: To construct and identify the RNAi lentiviral vector targeting rat DOT1L gene.
METHODS: DOT1L-specific shRNA sequence was designed, synthesized, and cloned into the lentivirus vector, and the recombinant lentivirus was obtained after packaging, amplification and purification. The interference effect on the expression of DOT1L gene in rat cells in primary rat vascular smooth muscle cells was identified by western blot assay.
RESULTS AND CONCLUSION: The rat DOT1L RNAi lentivirus vector was successfully constructed, and the final recombinant lentivirus titer was 8×108 PFU/mL. This vector can significantly reduce the protein level of DOT1L in vascular smooth muscle cells. The lentiviral vector can effectively silence the expression of DOT1L gene, which provides a reliable tool for further study of the biological function of DOT1L in rats.

Key words: rat, disruptor of telomeric silencing 1-like (DOT1L), RNA interference, lentiviral vector, plasmid, histone modification, cardiovascular disease, 293T human embryonic kidney cells

CLC Number:

Yang Zishu, Yang Jian, Huang Cuiyuan, Cheng Chen, Wu Di, Zhang Jing. Construction and identification of RNA interference lentiviral vector for rat disruptor of telomeric silencing 1-like gene[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(31): 8197-8204.

Figures/Tables 3

References

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Construction and identification of RNA interference lentiviral vector for rat disruptor of telomeric silencing 1-like gene

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