Overexpression of lncRNAs with endogenous lengths and functions using a lncRNA delivery system based on transposon
Abstract Background Long noncoding RNAs (lncRNAs) play important roles in many physiological and pathological processes, this indicates that lncRNAs can serve as potential targets for gene therapy. Stable expression is a fundamental technology in the study of lncRNAs. The lentivirus is one of the mo...
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Format: | Article |
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BMC
2021-10-01
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Series: | Journal of Nanobiotechnology |
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Online Access: | https://doi.org/10.1186/s12951-021-01044-7 |
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Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yin Zhang Yong-Xin Huang Xin Jin Jie Chen Li Peng Dan-Lan Wang Yun Li Xin-Yi Yao Jian-You Liao Jie-Hua He KaiShun Hu Daning Lu Yabin Guo Dong Yin |
spellingShingle |
Yin Zhang Yong-Xin Huang Xin Jin Jie Chen Li Peng Dan-Lan Wang Yun Li Xin-Yi Yao Jian-You Liao Jie-Hua He KaiShun Hu Daning Lu Yabin Guo Dong Yin Overexpression of lncRNAs with endogenous lengths and functions using a lncRNA delivery system based on transposon Journal of Nanobiotechnology lncRNA Non-viral delivery Ectopic overexpression Sleeping Beauty transposon Lentivirus |
author_facet |
Yin Zhang Yong-Xin Huang Xin Jin Jie Chen Li Peng Dan-Lan Wang Yun Li Xin-Yi Yao Jian-You Liao Jie-Hua He KaiShun Hu Daning Lu Yabin Guo Dong Yin |
author_sort |
Yin Zhang |
title |
Overexpression of lncRNAs with endogenous lengths and functions using a lncRNA delivery system based on transposon |
title_short |
Overexpression of lncRNAs with endogenous lengths and functions using a lncRNA delivery system based on transposon |
title_full |
Overexpression of lncRNAs with endogenous lengths and functions using a lncRNA delivery system based on transposon |
title_fullStr |
Overexpression of lncRNAs with endogenous lengths and functions using a lncRNA delivery system based on transposon |
title_full_unstemmed |
Overexpression of lncRNAs with endogenous lengths and functions using a lncRNA delivery system based on transposon |
title_sort |
overexpression of lncrnas with endogenous lengths and functions using a lncrna delivery system based on transposon |
publisher |
BMC |
series |
Journal of Nanobiotechnology |
issn |
1477-3155 |
publishDate |
2021-10-01 |
description |
Abstract Background Long noncoding RNAs (lncRNAs) play important roles in many physiological and pathological processes, this indicates that lncRNAs can serve as potential targets for gene therapy. Stable expression is a fundamental technology in the study of lncRNAs. The lentivirus is one of the most widely used delivery systems for stable expression. However, it was initially designed for mRNAs, and the applicability of lentiviral vectors for lncRNAs is largely unknown. Results We found that the lentiviral vector produces lncRNAs with improper termination, appending an extra fragment of ~ 2 kb to the 3ʹ-end. Consequently, the secondary structures were changed, the RNA–protein interactions were blocked, and the functions were impaired in certain lncRNAs, which indicated that lentiviral vectors are not ideal delivery systems of lncRNAs. Here, we developed a novel lncRNA delivery method called the Expression of LncRNAs with Endogenous Characteristics using the Transposon System (ELECTS). By inserting a termination signal after the lncRNA sequence, ELECTS produces transcripts without 3ʹ-flanking sequences and retains the native features and function of lncRNAs, which cannot be achieved by lentiviral vectors. Moreover, ELECTS presents no potential risk of infection for the operators and it takes much less time. ELECTS provides a reliable, convenient, safe, and efficient delivery method for stable expression of lncRNAs. Conclusions Our study demonstrated that improper transcriptional termination from lentiviral vectors have fundamental effects on molecular action and cellular function of lncRNAs. The ELECTS system developed in this study will provide a convenient and reliable method for the lncRNA study. Graphic Abstract |
topic |
lncRNA Non-viral delivery Ectopic overexpression Sleeping Beauty transposon Lentivirus |
url |
https://doi.org/10.1186/s12951-021-01044-7 |
work_keys_str_mv |
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doaj-5856d35e93ab4ba0ad1e8fb8f7c0dac72021-10-03T11:43:00ZengBMCJournal of Nanobiotechnology1477-31552021-10-0119111710.1186/s12951-021-01044-7Overexpression of lncRNAs with endogenous lengths and functions using a lncRNA delivery system based on transposonYin Zhang0Yong-Xin Huang1Xin Jin2Jie Chen3Li Peng4Dan-Lan Wang5Yun Li6Xin-Yi Yao7Jian-You Liao8Jie-Hua He9KaiShun Hu10Daning Lu11Yabin Guo12Dong Yin13Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Research Center of Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen UniversityAbstract Background Long noncoding RNAs (lncRNAs) play important roles in many physiological and pathological processes, this indicates that lncRNAs can serve as potential targets for gene therapy. Stable expression is a fundamental technology in the study of lncRNAs. The lentivirus is one of the most widely used delivery systems for stable expression. However, it was initially designed for mRNAs, and the applicability of lentiviral vectors for lncRNAs is largely unknown. Results We found that the lentiviral vector produces lncRNAs with improper termination, appending an extra fragment of ~ 2 kb to the 3ʹ-end. Consequently, the secondary structures were changed, the RNA–protein interactions were blocked, and the functions were impaired in certain lncRNAs, which indicated that lentiviral vectors are not ideal delivery systems of lncRNAs. Here, we developed a novel lncRNA delivery method called the Expression of LncRNAs with Endogenous Characteristics using the Transposon System (ELECTS). By inserting a termination signal after the lncRNA sequence, ELECTS produces transcripts without 3ʹ-flanking sequences and retains the native features and function of lncRNAs, which cannot be achieved by lentiviral vectors. Moreover, ELECTS presents no potential risk of infection for the operators and it takes much less time. ELECTS provides a reliable, convenient, safe, and efficient delivery method for stable expression of lncRNAs. Conclusions Our study demonstrated that improper transcriptional termination from lentiviral vectors have fundamental effects on molecular action and cellular function of lncRNAs. The ELECTS system developed in this study will provide a convenient and reliable method for the lncRNA study. Graphic Abstracthttps://doi.org/10.1186/s12951-021-01044-7lncRNANon-viral deliveryEctopic overexpressionSleeping Beauty transposonLentivirus |