Small Molecular Inhibitors of DNA Double Strand Break Repair Pathways Increase the ANTI-HBV Activity of CRISPR/Cas9
Kostyusheva A. P., Kostyushev D. S., Brezgin S. A., Zarifyan D. N., Volchkova E. V., Chulanov V. P.
Molecular Biology
Vol.53, Issue2, P. 274-285
Опубликовано: 2019
Тип ресурса: Статья
DOI:10.1134/S0026893319010072
Аннотация:
Abstract: The CRISPR/Cas9 nuclease system can effectively suppress the replication of the hepatitis B virus (HBV), while covalently closed circular DNA (cccDNA), a highly resistant form of the virus, persists in the nuclei of infected cells. The most common outcome of DNA double-strand breaks (DSBs) in cccDNA caused by CRISPR/Cas9 is double-strand break repair by nonhomologous end-joining, which results in insertion/deletion mutations. Modulation of the DNA double-strand break repair pathways by small molecules was shown to stimulate CRISPR/Cas9 activity and may potentially be utilized to enhance the elimination of HBV cccDNA. In this work, we used inhibitors of homologous (RI-1) and nonhomologous (NU7026) end-joining and their combination to stimulate antiviral activity of CRISPR/Cas9 on two cell models of HBV in vitro, i.e., the HepG2-1.1merHBV cells containing the HBV genome under the tet-on regulated cytomegalovirus promoter and the HepG2-1.5merHBV cells containing constitutive exp
Ключевые слова:
covalently closed circular DNA; CRISPR/Cas9; DNA double-strand breaks; hepatitis B virus; HR; lentiviral transduction; NHEJ
Язык текста: Английский
ISSN: 1608-3245
Kostyusheva A. P.
Kostyushev D. S.
Brezgin S. A.
Zarifyan D. N.
Volchkova E. V. Elena Vasilyevna 1953-
Chulanov V. P. Vladimir Petrovich 1970-
Костюшева А. П.
Костюшев Д. С.
Брезгин С. А.
Зарифян Д. Н.
Волчкова Е. В. Елена Васильевна 1953-
Чуланов В. П. Владимир Петрович 1970-
Small Molecular Inhibitors of DNA Double Strand Break Repair Pathways Increase the ANTI-HBV Activity of CRISPR/Cas9
Текст визуальный непосредственный
Molecular Biology
Pleiades Publishing, Inc.
Vol.53, Issue2 P. 274-285
2019
Статья
covalently closed circular DNA CRISPR/Cas9 DNA double-strand breaks hepatitis B virus HR lentiviral transduction NHEJ
Abstract: The CRISPR/Cas9 nuclease system can effectively suppress the replication of the hepatitis B virus (HBV), while covalently closed circular DNA (cccDNA), a highly resistant form of the virus, persists in the nuclei of infected cells. The most common outcome of DNA double-strand breaks (DSBs) in cccDNA caused by CRISPR/Cas9 is double-strand break repair by nonhomologous end-joining, which results in insertion/deletion mutations. Modulation of the DNA double-strand break repair pathways by small molecules was shown to stimulate CRISPR/Cas9 activity and may potentially be utilized to enhance the elimination of HBV cccDNA. In this work, we used inhibitors of homologous (RI-1) and nonhomologous (NU7026) end-joining and their combination to stimulate antiviral activity of CRISPR/Cas9 on two cell models of HBV in vitro, i.e., the HepG2-1.1merHBV cells containing the HBV genome under the tet-on regulated cytomegalovirus promoter and the HepG2-1.5merHBV cells containing constitutive exp