Cellular and Molecular Life Sciences, volume 76, issue 9, pages 1779-1794
Orthologous CRISPR/Cas9 systems for specific and efficient degradation of covalently closed circular DNA of hepatitis B virus
1
Publication type: Journal Article
Publication date: 2019-01-23
Journal:
Cellular and Molecular Life Sciences
Quartile SCImago
Q1
Quartile WOS
Q1
Impact factor: 8
ISSN: 1420682X, 14209071
Molecular Biology
Pharmacology
Cell Biology
Molecular Medicine
Cellular and Molecular Neuroscience
Abstract
Covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV) is the major cause of viral persistence and chronic hepatitis B. CRISPR/Cas9 nucleases can specifically target HBV cccDNA for decay, but off-target effects of nucleases in the human genome limit their clinical utility. CRISPR/Cas9 systems from four different species were co-expressed in cell lines with guide RNAs targeting conserved regions of the HBV genome. CRISPR/Cas9 systems from Streptococcus pyogenes (Sp) and Streptococcus thermophilus (St) targeting conserved regions of the HBV genome blocked HBV replication and, most importantly, resulted in degradation of over 90% of HBV cccDNA by 6 days post-transfection. Degradation of HBV cccDNA was impaired by inhibition of non-homologous end-joining pathway and resulted in an erroneous repair of HBV cccDNA. HBV cccDNA methylation also affected antiviral activity of CRISPR/Cas9. Single-nucleotide HBV genetic variants did not impact anti-HBV activity of St CRISPR/Cas9, suggesting its utility in targeting many HBV variants. However, two or more mismatches impaired or blocked CRISPR/Cas9 activity, indicating that host DNA will not likely be targeted. Deep sequencing revealed that Sp CRISPR/Cas9 induced off-target mutagenesis, whereas St CRISPR/Cas9 had no effect on the host genome. St CRISPR/Cas9 system represents the safest system with high anti-HBV activity.
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Citations by publishers
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6 publications, 11.54%
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- We do not take into account publications that without a DOI.
- Statistics recalculated only for publications connected to researchers, organizations and labs registered on the platform.
- Statistics recalculated weekly.
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Kostyushev D. et al. Orthologous CRISPR/Cas9 systems for specific and efficient degradation of covalently closed circular DNA of hepatitis B virus // Cellular and Molecular Life Sciences. 2019. Vol. 76. No. 9. pp. 1779-1794.
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Kostyushev D., Brezgin S., Kostyusheva A., Zarifyan D., Goptar I., Chulanov V. Orthologous CRISPR/Cas9 systems for specific and efficient degradation of covalently closed circular DNA of hepatitis B virus // Cellular and Molecular Life Sciences. 2019. Vol. 76. No. 9. pp. 1779-1794.
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TY - JOUR
DO - 10.1007/s00018-019-03021-8
UR - https://doi.org/10.1007%2Fs00018-019-03021-8
TI - Orthologous CRISPR/Cas9 systems for specific and efficient degradation of covalently closed circular DNA of hepatitis B virus
T2 - Cellular and Molecular Life Sciences
AU - Kostyushev, Dmitry
AU - Kostyusheva, Anastasiya
AU - Zarifyan, Dmitry
AU - Goptar, Irina
AU - Chulanov, Vladimir
AU - Brezgin, Sergey
PY - 2019
DA - 2019/01/23 00:00:00
PB - Springer Nature
SP - 1779-1794
IS - 9
VL - 76
SN - 1420-682X
SN - 1420-9071
ER -
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@article{2019_Kostyushev,
author = {Dmitry Kostyushev and Anastasiya Kostyusheva and Dmitry Zarifyan and Irina Goptar and Vladimir Chulanov and Sergey Brezgin},
title = {Orthologous CRISPR/Cas9 systems for specific and efficient degradation of covalently closed circular DNA of hepatitis B virus},
journal = {Cellular and Molecular Life Sciences},
year = {2019},
volume = {76},
publisher = {Springer Nature},
month = {jan},
url = {https://doi.org/10.1007%2Fs00018-019-03021-8},
number = {9},
pages = {1779--1794},
doi = {10.1007/s00018-019-03021-8}
}
Cite this
MLA
Copy
Kostyushev, Dmitry, et al. “Orthologous CRISPR/Cas9 systems for specific and efficient degradation of covalently closed circular DNA of hepatitis B virus.” Cellular and Molecular Life Sciences, vol. 76, no. 9, Jan. 2019, pp. 1779-1794. https://doi.org/10.1007%2Fs00018-019-03021-8.