Hepatitis B virus-like particle: targeted delivery of plasmid expressing short hairpin RNA for silencing the Bcl-2 gene in cervical cancer cells

Gene therapy research has advanced to clinical trials, but it is hampered by unstable nucleic acids packaged inside carriers and there is a lack of specificity towards targeted sites in the body. This study aims to address gene therapy limitations by encapsidating a plasmid synthesizing a short hair...

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Main Authors: Akwiditya, Made Angga, Yong, Chean Yeah, Yusof, Mohd Termizi, Abdul Razak, Mariatulqabtiah, Ho, Kok Lian, Tan, Wen Siang
Format: Article
Language:English
Published: Multidisciplinary Digital Publishing Institute 2021
Online Access:http://psasir.upm.edu.my/id/eprint/97514/1/ABSTRACT.pdf
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author Akwiditya, Made Angga
Yong, Chean Yeah
Yusof, Mohd Termizi
Abdul Razak, Mariatulqabtiah
Ho, Kok Lian
Tan, Wen Siang
author_facet Akwiditya, Made Angga
Yong, Chean Yeah
Yusof, Mohd Termizi
Abdul Razak, Mariatulqabtiah
Ho, Kok Lian
Tan, Wen Siang
author_sort Akwiditya, Made Angga
collection UPM
description Gene therapy research has advanced to clinical trials, but it is hampered by unstable nucleic acids packaged inside carriers and there is a lack of specificity towards targeted sites in the body. This study aims to address gene therapy limitations by encapsidating a plasmid synthesizing a short hairpin RNA (shRNA) that targets the anti-apoptotic Bcl-2 gene using truncated hepatitis B core antigen (tHBcAg) virus-like particle (VLP). A shRNA sequence targeting anti-apoptotic Bcl-2 was synthesized and cloned into the pSilencer 2.0-U6 vector. The recombinant plasmid, namely PshRNA, was encapsidated inside tHBcAg VLP and conjugated with folic acid (FA) to produce FA-tHBcAg-PshRNA VLP. Electron microscopy revealed that the FA-tHBcAg-PshRNA VLP has an icosahedral structure that is similar to the unmodified tHBcAg VLP. Delivery of FA-tHBcAg-PshRNA VLP into HeLa cells overexpressing the folate receptor significantly down regulated the expression of anti-apoptotic Bcl-2 at 48 and 72 h post-transfection. The 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay demonstrated that the cells’ viability was significantly reduced from 89.46% at 24 h to 64.52% and 60.63%, respectively, at 48 and 72 h post-transfection. As a conclusion, tHBcAg VLP can be used as a carrier for a receptor-mediated targeted delivery of a therapeutic plasmid encoding shRNA for gene silencing in cancer cells.
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spelling upm.eprints-975142022-07-27T02:48:39Z http://psasir.upm.edu.my/id/eprint/97514/ Hepatitis B virus-like particle: targeted delivery of plasmid expressing short hairpin RNA for silencing the Bcl-2 gene in cervical cancer cells Akwiditya, Made Angga Yong, Chean Yeah Yusof, Mohd Termizi Abdul Razak, Mariatulqabtiah Ho, Kok Lian Tan, Wen Siang Gene therapy research has advanced to clinical trials, but it is hampered by unstable nucleic acids packaged inside carriers and there is a lack of specificity towards targeted sites in the body. This study aims to address gene therapy limitations by encapsidating a plasmid synthesizing a short hairpin RNA (shRNA) that targets the anti-apoptotic Bcl-2 gene using truncated hepatitis B core antigen (tHBcAg) virus-like particle (VLP). A shRNA sequence targeting anti-apoptotic Bcl-2 was synthesized and cloned into the pSilencer 2.0-U6 vector. The recombinant plasmid, namely PshRNA, was encapsidated inside tHBcAg VLP and conjugated with folic acid (FA) to produce FA-tHBcAg-PshRNA VLP. Electron microscopy revealed that the FA-tHBcAg-PshRNA VLP has an icosahedral structure that is similar to the unmodified tHBcAg VLP. Delivery of FA-tHBcAg-PshRNA VLP into HeLa cells overexpressing the folate receptor significantly down regulated the expression of anti-apoptotic Bcl-2 at 48 and 72 h post-transfection. The 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay demonstrated that the cells’ viability was significantly reduced from 89.46% at 24 h to 64.52% and 60.63%, respectively, at 48 and 72 h post-transfection. As a conclusion, tHBcAg VLP can be used as a carrier for a receptor-mediated targeted delivery of a therapeutic plasmid encoding shRNA for gene silencing in cancer cells. Multidisciplinary Digital Publishing Institute 2021 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/97514/1/ABSTRACT.pdf Akwiditya, Made Angga and Yong, Chean Yeah and Yusof, Mohd Termizi and Abdul Razak, Mariatulqabtiah and Ho, Kok Lian and Tan, Wen Siang (2021) Hepatitis B virus-like particle: targeted delivery of plasmid expressing short hairpin RNA for silencing the Bcl-2 gene in cervical cancer cells. International Journal of Molecular Sciences, 22 (5). art. no. 2320. pp. 1-14. ISSN 1422-0067 https://www.mdpi.com/1422-0067/22/5/2320?type=check_update&version=2 10.3390/ijms22052320
spellingShingle Akwiditya, Made Angga
Yong, Chean Yeah
Yusof, Mohd Termizi
Abdul Razak, Mariatulqabtiah
Ho, Kok Lian
Tan, Wen Siang
Hepatitis B virus-like particle: targeted delivery of plasmid expressing short hairpin RNA for silencing the Bcl-2 gene in cervical cancer cells
title Hepatitis B virus-like particle: targeted delivery of plasmid expressing short hairpin RNA for silencing the Bcl-2 gene in cervical cancer cells
title_full Hepatitis B virus-like particle: targeted delivery of plasmid expressing short hairpin RNA for silencing the Bcl-2 gene in cervical cancer cells
title_fullStr Hepatitis B virus-like particle: targeted delivery of plasmid expressing short hairpin RNA for silencing the Bcl-2 gene in cervical cancer cells
title_full_unstemmed Hepatitis B virus-like particle: targeted delivery of plasmid expressing short hairpin RNA for silencing the Bcl-2 gene in cervical cancer cells
title_short Hepatitis B virus-like particle: targeted delivery of plasmid expressing short hairpin RNA for silencing the Bcl-2 gene in cervical cancer cells
title_sort hepatitis b virus like particle targeted delivery of plasmid expressing short hairpin rna for silencing the bcl 2 gene in cervical cancer cells
url http://psasir.upm.edu.my/id/eprint/97514/1/ABSTRACT.pdf
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