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TALEN based HPV-E7 editing triggers necrotic cell death in cervical cancer cells

Human Papillomavirus E7 and E6 oncoproteins have been considered as suitable candidate anti-viral targets since they cause malignant conversion in cervical cancers. Transcription Activator-Like Effector Nucleases (TALENs) are recent editing tools to knockout genes by inducing double stranded breaks...

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Autores principales: Shankar, Sumitra, Prasad, Deepti, Sanawar, Rahul, Das, Ani V., Pillai, M. Radhakrishna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5511212/
https://www.ncbi.nlm.nih.gov/pubmed/28710417
http://dx.doi.org/10.1038/s41598-017-05696-0
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author Shankar, Sumitra
Prasad, Deepti
Sanawar, Rahul
Das, Ani V.
Pillai, M. Radhakrishna
author_facet Shankar, Sumitra
Prasad, Deepti
Sanawar, Rahul
Das, Ani V.
Pillai, M. Radhakrishna
author_sort Shankar, Sumitra
collection PubMed
description Human Papillomavirus E7 and E6 oncoproteins have been considered as suitable candidate anti-viral targets since they cause malignant conversion in cervical cancers. Transcription Activator-Like Effector Nucleases (TALENs) are recent editing tools to knockout genes by inducing double stranded breaks at specific sites in the genome. In here, we have designed specific TALENs to target E7 and analyzed their efficiency in inducing cell death in cervical cancer cells. We found that designed TALENs could yield about 10–12% editing activity as observed from T7E1 and nuclease resistance assays. Down-regulation of E7 and E6 was further evident at the transcript as well as proteins levels indicating that the selected TALENs were effective. TALEN-mediated E7 editing led to cell death as ascertained by cell cycle and Annexin V assays. Annexin profiling suggested that cell death could be due to necrosis as observed by upregulation of necrotic markers such as LDH A, Rip-1, and Cyclophilin A. Necrosis appears to be a better therapeutic response as it could further activate pro-inflammatory cytokines to attract immune cells to eliminate HPV-integrated cells and therefore TALEN editing strategy has the potential to be a promising tool as an adjuvant therapy in cervical cancer along with surgery.
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spelling pubmed-55112122017-07-17 TALEN based HPV-E7 editing triggers necrotic cell death in cervical cancer cells Shankar, Sumitra Prasad, Deepti Sanawar, Rahul Das, Ani V. Pillai, M. Radhakrishna Sci Rep Article Human Papillomavirus E7 and E6 oncoproteins have been considered as suitable candidate anti-viral targets since they cause malignant conversion in cervical cancers. Transcription Activator-Like Effector Nucleases (TALENs) are recent editing tools to knockout genes by inducing double stranded breaks at specific sites in the genome. In here, we have designed specific TALENs to target E7 and analyzed their efficiency in inducing cell death in cervical cancer cells. We found that designed TALENs could yield about 10–12% editing activity as observed from T7E1 and nuclease resistance assays. Down-regulation of E7 and E6 was further evident at the transcript as well as proteins levels indicating that the selected TALENs were effective. TALEN-mediated E7 editing led to cell death as ascertained by cell cycle and Annexin V assays. Annexin profiling suggested that cell death could be due to necrosis as observed by upregulation of necrotic markers such as LDH A, Rip-1, and Cyclophilin A. Necrosis appears to be a better therapeutic response as it could further activate pro-inflammatory cytokines to attract immune cells to eliminate HPV-integrated cells and therefore TALEN editing strategy has the potential to be a promising tool as an adjuvant therapy in cervical cancer along with surgery. Nature Publishing Group UK 2017-07-14 /pmc/articles/PMC5511212/ /pubmed/28710417 http://dx.doi.org/10.1038/s41598-017-05696-0 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Shankar, Sumitra
Prasad, Deepti
Sanawar, Rahul
Das, Ani V.
Pillai, M. Radhakrishna
TALEN based HPV-E7 editing triggers necrotic cell death in cervical cancer cells
title TALEN based HPV-E7 editing triggers necrotic cell death in cervical cancer cells
title_full TALEN based HPV-E7 editing triggers necrotic cell death in cervical cancer cells
title_fullStr TALEN based HPV-E7 editing triggers necrotic cell death in cervical cancer cells
title_full_unstemmed TALEN based HPV-E7 editing triggers necrotic cell death in cervical cancer cells
title_short TALEN based HPV-E7 editing triggers necrotic cell death in cervical cancer cells
title_sort talen based hpv-e7 editing triggers necrotic cell death in cervical cancer cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5511212/
https://www.ncbi.nlm.nih.gov/pubmed/28710417
http://dx.doi.org/10.1038/s41598-017-05696-0
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