actabiomedicaActa Biomedica ScientificaActa Biomedica Scientifica2541-94202587-9596Scientific Centre for Family Health and Human Reproduction Problems10.29413/ABS.2023-8.1.5actabiomedica-3986Research ArticleГЕНЕТИКА, ПРОТЕОМИКА И МЕТАБОЛОМИКАGENETICS, PROTEOMICS AND METABOLOMICSПерспективы применения системы CRISPR-Cas9 в лечении вирусных заболеваний человекаProspects for using CRISPR-Cas9 system in the treatment of human viral diseaseshttps://orcid.org/0000-0001-5474-1080ЗиганшинА. М.ZiganshinA. M.

Зиганшин Айдар Миндиярович – кандидат медицинских наук, доцент кафедры акушерства и гинекологии с курсом ИДПО, 

450008, г. Уфа, ул. Ленина, 3

Aydar M. Ziganshin – Cand. Sc. (Med.), Associate Professor at the Department of Obstetrics and Gynecology with the course of the Institute of Advanced Professional Education, 

Lenina str. 3, Ufa 450008

zigaidar@yandex.ruhttps://orcid.org/0000-0001-7490-3710МулюковА. Р.MulyukovA. R.

Мулюков Айрат Рамильевич – студент, 

450008, г. Уфа, ул. Ленина, 3

Airat R. Mulykov – Student, 

Lenina str. 3, Ufa 450008

mulykov.165@mail.ruhttps://orcid.org/0000-0003-1061-3006ОмаровМ. А.OmarovM. A.

Омаров Магомед Абдурахманович – студент, 

450008, г. Уфа, ул. Ленина, 3

Magomed A. Omarov – Student,

Lenina str. 3, Ufa 450008

mmaaggaa_omarov@mail.ruhttps://orcid.org/0000-0002-5961-5400МудровВ. А.MudrovV. A.

Мудров Виктор Андреевич – кандидат медицинских наук, доцент кафедры акушерства и гинекологии лечебного и стоматологического факультетов, 

672000, г. Чита, ул. Горького, 39а

Viktor  A. Mudrov – Cand.  Sc. (Med.), Associate Professor at the Department of Obstetrics and Gynecology of the Faculties of Medicine and Dentistry,

Gorkogo str. 39A, Chita 672000

mudrov_viktor@mail.ruhttps://orcid.org/0000-0002-2629-2051ХалитоваР. Ш.KhalitovaR. Sh.

Халитова Регина Шамилевна – аспирант кафедры акушерства и гинекологии с курсом ИДПО, 

450008, г. Уфа, ул. Ленина, 3

Regina Sh. Khalitova – Postgraduate at the Department of Obstetrics and Gynecology with the course of the Institute of Advanced Professional Education,

Lenina str. 3, Ufa 450008

regina_badranova@mail.ruФГБОУ ВО «Башкирский государственный медицинский университет» Минздрава РоссииBashkir State Medical UniversityФГБОУ ВО «Читинская государственная медицинская академия» Минздрава РоссииChita State Medical Academy202307032023814050Copyright © Зиганшин А.М., Мулюков А.Р., Омаров М.А., Мудров В.А., Халитова Р.Ш., 20232023Зиганшин А.М., Мулюков А.Р., Омаров М.А., Мудров В.А., Халитова Р.Ш.Ziganshin A.M., Mulyukov A.R., Omarov M.A., Mudrov V.A., Khalitova R.S.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.actabiomedica.ru/jour/article/view/3986

Цель данной статьи – провести анализ возможности применения генетических механизмов технологии CRISPR-Cas9 в профилактике и лечении некоторых вирусных заболеваний.

Материалы и методы

Материалы и методы. Поиск публикаций проводился в российской и зарубежной литературе в следующих поисковых системах: РИНЦ, Сyberleninka, eLibrary, PubMed, библиотека Cochrane и др. Проведён обзор отечественных и международных научных работ по теме исследования с использованием поисковых ключевых слов: CRISPR, генная инженерия, редактирование генома, Cas9, sgRNA.

Результаты

Результаты. Проведён обзор использования метода CRISPR-Cas9 («генетических ножниц») в качестве генной терапии некоторых вирусных заболеваний, раскрыты его основные преимущества и недостатки. Анализ данных научных исследований в сфере генетических методов исследования за последнее десятилетие раскрывает основные аспекты технологии CRISPR-Cas9, современную классификацию и перспективы применения данной технологии в клинической практике с целью терапии и профилактики вирусных заболеваний человека. Рассматриваются возможности создания более многофункциональной и стабильной версии технологии CRISPR-Cas9. Особое внимание уделяется технологическим сложностям и препятствиям, которые встают перед учёными при внедрении данной системы для целевого применения в клинической медицине.

Заключение

Заключение. Открытие «генетических ножниц» произвело революцию во всей медицине. Перед медицинским сообществом открылись широкие возможности для создания новых методов лечения множества вирусных заболеваний и создания условий для ранней профилактики. В перспективе при внедрении данной технологии в клиническую практику станет возможной терапия нозологий, ранее не отвечавших на проводимую терапию и считавшихся неизлечимыми. 

The aim

The aim. To analyze the possibility of using the genetic mechanisms of CRISPR-Cas9 technology in the prevention and treatment of certain viral diseases.

Materials and methods

Materials and methods. The search for publications was carried out in Russian and foreign literature using the following search engines: RSCI, Cyberleninka, eLibrary, PubMed, Cochrane Library, etc. A review of domestic and international scientific papers on the research topic was carried out using search keywords: CRISPR, genetic engineering, genome editing, Cas9, sgRNA.

Results

Results. A review of using CRISPR-Cas9 method (“genetic scissors”) as a gene therapy for some viral diseases was carried out, and its main advantages and disadvantages were revealed. An analysis of the data of scientific studies on genetic research methods over the past decade discovers the main aspects of CRISPR-Cas9 technology, modern classification and prospects for using this technology in clinical practice for the treatment and prevention of human viral diseases. The possibilities of creating a more versatile and stable version of the CRISPR-Cas9 technology are considered. Particular attention is paid to the technological difficulties and obstacles that scientists face when implementing this system for targeted use in clinical medicine.

Conclusion

Conclusion. One of the rapidly developing areas in science giving promising prospects for modern healthcare is genetic engineering, especially in cases where scientific developments are applied in clinical practice. The discovery of “genetic scissors” technology has revolutionized all medicine. Wide opportunities for developing new treatment methods for many viral diseases and creating conditions for their early prevention opened up for the medical community. In the future, with the introduction of this technology into clinical practice, it will become possible to treat diseases that have not previously responded to ongoing therapy and were considered incurable. 

CRISPRгенная инженерияредактирование геномаCas9sgRNAвирусные заболеванияинфекцииCRISPRgenetic engineeringgenome editingCas9sgRNAviral diseasesinfectionsReferencesЗиганшин А.М., Мулюков А.Р. Механизмы иммунопатологии сепсиса вирусной этиологии при COVID-19. Сибирское медицинское обозрение. 2021; (6): 35-43. doi: 10.20333/25000136-2021-6-35-43Ziganshin AM, Mulykov AR. Immunopathological mechanisms in sepsis of viral etiology in COVID-19. Siberian Medical Review. 2021; (6): 35-43. (In Russ.). doi: 10.20333/25000136-2021-6-35-43Bellizzi A, Ahye N, Jalagadugula G, Wollebo HS. A broad application of CRISPR Cas9 in infectious diseases of central nervous system. J Neuroimmune Pharmacol. 2019; 14(4): 578-594. doi: 10.1007/s11481-019-09878-7Bellizzi A, Ahye N, Jalagadugula G, Wollebo HS. A broad application of CRISPR Cas9 in infectious diseases of central nervous system. J Neuroimmune Pharmacol. 2019; 14(4): 578-594. doi: 10.1007/s11481-019-09878-7Ishino Y, Krupovic M, Forterre P. History of CRISPR-Cas from encounter with a mysterious repeated sequence to genome editing technology. J Bacteriol. 2018; 200(7): e00580-17. doi: 10.1128/JB.00580-17Ishino Y, Krupovic M, Forterre P. History of CRISPR-Cas from encounter with a mysterious repeated sequence to genome editing technology. J Bacteriol. 2018; 200(7): e00580-17. doi: 10.1128/ JB.00580-17Makarova KS, Grishin NV, Shabalina SA, Wolf YI, Koonin EV. A putative RNA-interference-based immune system in prokaryotes: Computational analysis of the predicted enzymatic machinery, functional analogies with eukaryotic RNAi, and hypothetical mechanisms of action. Biol Direct. 2006; 1: 7. doi: 10.1186/1745-6150-1-7Makarova KS, Grishin NV, Shabalina SA, Wolf YI, Koonin EV. A putative RNA-interference-based immune system in prokaryotes: Computational analysis of the predicted enzymatic machinery, functional analogies with eukaryotic RNAi, and hypothetical mechanisms of action. Biol Direct. 2006; 1: 7. doi: 10.1186/1745-6150-1-7Barrangou R, Doudna JA. Applications of CRISPR technologies in research and beyond. Nature Biotechnol. 2016; 34(9): 933- 941. doi: 10.1038/nbt.3659Barrangou R, Doudna JA. Applications of CRISPR technologies in research and beyond. Nature Biotechnol. 2016; 34(9): 933- 941. doi: 10.1038/nbt.3659Bollen Y, Post J, Koo BK, Snippert HJG. How to create state-of-the-art genetic model systems: Strategies for optimal CRISPR-mediated genome editing. Nucleic Acids Res. 2018; 46(13): 6435-6454. doi: 10.1093/nar/gky571Bollen Y, Post J, Koo BK, Snippert HJG. How to create state-of-the-art genetic model systems: Strategies for optimal CRISPR-mediated genome editing. Nucleic Acids Res. 2018; 46(13): 6435-6454. doi: 10.1093/nar/gky571Zhuo C, Zhang J, Lee JH, Jiao J, Cheng D, Liu L, et al. Spatiotemporal control of CRISPR/Cas9 gene editing. Signal Transduct Target Ther. 2021; 6(1): 238. doi: 10.1038/s41392-021-00645-wZhuo C, Zhang J, Lee JH, Jiao J, Cheng D, Liu L, et al. Spatiotemporal control of CRISPR/Cas9 gene editing. Signal Transduct Target Ther. 2021; 6(1): 238. doi: 10.1038/s41392-021-00645-wChen S, Lee B, Lee AY, Modzelewski AJ, He L. Highly efficient mouse genome editing by CRISPR ribonucleoprotein electroporation of zygotes. J Biol Chem. 2016; 291(28): 14457-14467. doi: 10.1074/jbc.M116.733154Chen S, Lee B, Lee AY, Modzelewski AJ, He L. Highly efficient mouse genome editing by CRISPR ribonucleoprotein electroporation of zygotes. J Biol Chem. 2016; 291(28): 14457-14467. doi: 10.1074/jbc.M116.733154Tong S, Moyo B, Lee CM, Leong K, Bao G. Engineered materials for in vivo delivery of genome-editing machinery. Nat Rev Mater. 2019; 4: 726-737. doi: 10.1038/s41578-019-0145-9Tong S, Moyo B, Lee CM, Leong K, Bao G. Engineered materials for in vivo delivery of genome-editing machinery. Nat Rev Mater. 2019; 4: 726-737. doi: 10.1038/s41578-019-0145-9D’Agostino Y, D’Aniello S. Molecular basis, applications and challenges of CRISPR/Cas9: A continuously evolving tool for genome editing. Brief Funct Genomics. 2017; 16(4): 211-216. doi: 10.1093/bfgp/elw038D’Agostino Y, D’Aniello S. Molecular basis, applications and challenges of CRISPR/Cas9: A continuously evolving tool for genome editing. Brief Funct Genomics. 2017; 16(4): 211-216. doi: 10.1093/bfgp/elw038Chu VT, Weber T, Wefers B, Wurst W, Sander S, Rajewsky K, et al. Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nature Biotechnol. 2015; 33(5): 543-548. doi: 10.1038/nbt.3198Chu VT, Weber T, Wefers B, Wurst W, Sander S, Rajewsky K, et al. Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Nature Biotechnol. 2015; 33(5): 543-548. doi: 10.1038/nbt.3198Makarova KS, Wolf YI, Iranzo J, Shmakov SA, Alkhnbashi OS, Brouns SJJ, et al. Evolutionary classification of CRISPR-Cas systems: A burst of class 2 and derived variants. Nat Rev Microbiol. 2020; 18(2): 67-83. doi: 10.1038/s41579-019-0299-xMakarova KS, Wolf YI, Iranzo J, Shmakov SA, Alkhnbashi OS, Brouns SJJ, et al. Evolutionary classification of CRISPR-Cas systems: A burst of class 2 and derived variants. Nat Rev Microbiol. 2020; 18(2): 67-83. doi: 10.1038/s41579-019-0299-xChen JS, Dagdas YS, Kleinstiver BP, Welch MM, Sousa AA, Harrington LB, et al. Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017; 550(7676): 407-410. doi: 10.1038/nature24268Chen JS, Dagdas YS, Kleinstiver BP, Welch MM, Sousa AA, Harrington LB, et al. Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017; 550(7676): 407-410. doi: 10.1038/nature24268Crystal RG. Adenovirus: The first effective in vivo gene delivery vector. Hum Gene Ther. 2014; 25(1): 3-11. doi: 10.1089/hum.2013.2527Crystal RG. Adenovirus: The first effective in vivo gene delivery vector. Hum Gene Ther. 2014; 25(1): 3-11. doi: 10.1089/hum.2013.2527Charlesworth CT, Deshpande PS, Dever DP, Camarena J, Lemgart VT, Cromer MK, et al. Identification of preexisting adaptive immunity to Cas9 proteins in humans. Nature Med. 2019; 25(2): 249-254. doi: 10.1038/s41591-018-0326-xCharlesworth CT, Deshpande PS, Dever DP, Camarena J, Lemgart VT, Cromer MK, et al. Identification of preexisting adaptive immunity to Cas9 proteins in humans. Nature Med. 2019; 25(2): 249-254. doi: 10.1038/s41591-018-0326-xFu Y, Foden JA, Khayter C, Maeder ML, Reyon D, Joung JK, et al. High-frequency off-target mutagenesis induced by CRISPRCas nucleases in human cells. Nat Biotechnol. 2013; 31(9): 822-826. doi: 10.1038/nbt.2623Fu Y, Foden JA, Khayter C, Maeder ML, Reyon D, Joung JK, et al. High-frequency off-target mutagenesis induced by CRISPRCas nucleases in human cells. Nat Biotechnol. 2013; 31(9): 822-826. doi: 10.1038/nbt.2623Maartens G, Celum C, Lewin SR. HIV infection: Epidemiology, pathogenesis, treatment, and prevention. Lancet. 2014; 384(9939): 258-271. doi: 10.1016/S0140-6736(14)60164-1Maartens G, Celum C, Lewin SR. HIV infection: Epidemiology, pathogenesis, treatment, and prevention. Lancet. 2014; 384(9939): 258-271. doi: 10.1016/S0140-6736(14)60164-1Bialek JK, Dunay GA, Voges M, Schäfer C, Spohn M, Stucka R, et al. Targeted HIV-1 latency reversal using CRISPR/Cas9- derived transcriptional activator systems. PLoS One. 2016; 11(6): e0158294. doi: 10.1371/journal.pone.0158294Bialek JK, Dunay GA, Voges M, Schäfer C, Spohn M, Stucka R, et al. Targeted HIV-1 latency reversal using CRISPR/Cas9- derived transcriptional activator systems. PLoS One. 2016; 11(6): e0158294. doi: 10.1371/journal.pone.0158294Ebina H, Misawa N, Kanemura Y, Koyanagi Y. Harnessing the CRISPR/Cas9 system to disrupt latent HIV-1 provirus. Sci Rep. 2013; 3: 2510. doi: 10.1038/srep02510Ebina H, Misawa N, Kanemura Y, Koyanagi Y. Harnessing the CRISPR/Cas9 system to disrupt latent HIV-1 provirus. Sci Rep. 2013; 3: 2510. doi: 10.1038/srep02510Darcis G, Das AT, Berkhout B. Tackling HIV persistence: Pharmacological versus CRISPR-based shock strategies. Viruses. 2018; 10(4): 157. doi: 10.3390/v10040157Darcis G, Das AT, Berkhout B. Tackling HIV persistence: Pharmacological versus CRISPR-based shock strategies. Viruses. 2018; 10(4): 157. doi: 10.3390/v10040157Dash PK, Kaminski R, Bella R, Su H, Mathews S, Ahooyi TM, et al. Sequential LASER ART and CRISPR treatments eliminate HIV-1 in a subset of infected humanized mice. Nat Commun. 2019; 10(1): 2753. doi: 10.1038/s41467-019-10366-yDash PK, Kaminski R, Bella R, Su H, Mathews S, Ahooyi TM, et al. Sequential LASER ART and CRISPR treatments eliminate HIV-1 in a subset of infected humanized mice. Nat Commun. 2019; 10(1): 2753. doi: 10.1038/s41467-019-10366-yBella R, Kaminski R, Mancuso P, Young WB, Chen C, Sariyer R, et al. Removal of HIV DNA by CRISPR from patient blood engrafts in humanized mice. Mol Ther Nucleic Acids. 2018; 12: 275- 282. doi: 10.1016/j.omtn.2018.05.021Bella R, Kaminski R, Mancuso P, Young WB, Chen C, Sariyer R, et al. Removal of HIV DNA by CRISPR from patient blood engrafts in humanized mice. Mol Ther Nucleic Acids. 2018; 12: 275- 282. doi: 10.1016/j.omtn.2018.05.021Cheng R, Peng J, Yan Y, Cao P, Wang J, Qiu C, et al. Efficient gene editing in adult mouse livers via adenoviral delivery of CRISPR/Cas9. FEBS Lett. 2014; 588(21): 3954-3958. doi: 10.1016/j.febslet.2014.09.008Cheng R, Peng J, Yan Y, Cao P, Wang J, Qiu C, et al. Efficient gene editing in adult mouse livers via adenoviral delivery of CRISPR/Cas9. FEBS Lett. 2014; 588(21): 3954-3958. doi: 10.1016/j.febslet.2014.09.008Herrera-Carrillo E, Berkhout B. Attacking HIV-1 RNA versus DNA by sequence-specific approaches: RNAi versus CRISPR-Cas. Biochem Soc Trans. 2016; 44(5): 1355-1365. doi: 10.1042/BST20160060Herrera-Carrillo E, Berkhout B. Attacking HIV-1 RNA versus DNA by sequence-specific approaches: RNAi versus CRISPRCas. Biochem Soc Trans. 2016; 44(5): 1355-1365. doi: 10.1042/BST20160060Badia R, Ballana E, Castellví M, García-Vidal E, Pujantell M, Clotet B, et al. CD32 expression is associated to T-cell activation and is not a marker of the HIV-1 reservoir. Nature Communications. 2018; 9(1): 2739. doi: 10.1038/s41467-018-05157-wBadia R, Ballana E, Castellví M, García-Vidal E, Pujantell M, Clotet B, et al. CD32 expression is associated to T-cell activation and is not a marker of the HIV-1 reservoir. Nature Communications. 2018; 9(1): 2739. doi: 10.1038/s41467-018-05157-wDufour C, Claudel A, Joubarne N, Merindol N, Maisonnet T, Masroori N, et al. Editing of the human TRIM5 gene to introduce mutations with the potential to inhibit HIV-1. PLoS One. 2018; 13(1): e0191709. doi: 10.1371/journal.pone.0191709Dufour C, Claudel A, Joubarne N, Merindol N, Maisonnet T, Masroori N, et al. Editing of the human TRIM5 gene to introduce mutations with the potential to inhibit HIV-1. PLoS One. 2018; 13(1): e0191709. doi: 10.1371/journal.pone.0191709Bogerd HP, Kornepati AV, Marshall JB, Kennedy EM, Cullen BR. Specific induction of endogenous viral restriction factors using CRISPR/Cas-derived transcriptional activators. Proc Natl Acad Sci U S A. 2015; 112(52): E7249- E7256. doi: 10.1073/pnas.1516305112Bogerd HP, Kornepati AV, Marshall JB, Kennedy EM, Cullen BR. Specific induction of endogenous viral restriction factors using CRISPR/Cas-derived transcriptional activators. Proc Natl Acad Sci U S A. 2015; 112(52): E7249- E7256. doi: 10.1073/pnas.1516305112Chou YY, Krupp A, Kaynor C, Gaudin R, Ma M, Cahir-McFarland E, et al. Inhibition of JCPyV infection mediated by targeted viral genome editing using CRISPR/Cas9. Sci Rep. 2016; 6: 36921. doi: 10.1038/srep36921Chou YY, Krupp A, Kaynor C, Gaudin R, Ma M, Cahir-McFarland E, et al. Inhibition of JCPyV infection mediated by targeted viral genome editing using CRISPR/Cas9. Sci Rep. 2016; 6: 36921. doi: 10.1038/srep36921Wollebo HS, Bellizzi A, Kaminski R, Hu W, White MK, Khalili K. CRISPR/Cas9 system as an agent for eliminating polyomavirus JC infection. PLoS One. 2015; 10(9): e0136046. doi: 10.1371/journal.pone.0136046Wollebo HS, Bellizzi A, Kaminski R, Hu W, White MK, Khalili K. CRISPR/Cas9 system as an agent for eliminating polyomavirus JC infection. PLoS One. 2015; 10(9): e0136046. doi: 10.1371/journal.pone.0136046Bloom K, Maepa MB, Ely A, Arbuthnot P. Gene therapy for chronic HBV – Can we eliminate cccDNA? Genes (Basel). 2018; 9(4): 207. doi: 10.3390/genes9040207Bloom K, Maepa MB, Ely A, Arbuthnot P. Gene therapy for chronic HBV – Can we eliminate cccDNA? Genes (Basel). 2018; 9(4): 207. doi: 10.3390/genes9040207Chang J, Guo JT. Treatment of chronic hepatitis B with pattern recognition receptor agonists: Current status and potential for a cure. Antiviral Res. 2015; 121: 152-159. doi: 10.1016/j.antiviral.2015.07.006Chang J, Guo JT. Treatment of chronic hepatitis B with pattern recognition receptor agonists: Current status and potential for a cure. Antiviral Res. 2015; 121: 152-159. doi: 10.1016/j.antiviral.2015.07.006Dong C, Qu L, Wang H, Wei L, Dong Y, Xiong S. Targeting hepatitis B virus cccDNA by CRISPR/Cas9 nuclease efficiently inhibits viral replication. Antiviral Res. 2015; 118: 110-117. doi: 10.1016/j.antiviral.2015.03.015Dong C, Qu L, Wang H, Wei L, Dong Y, Xiong S. Targeting hepatitis B virus cccDNA by CRISPR/Cas9 nuclease efficiently inhibits viral replication. Antiviral Res. 2015; 118: 110-117. doi: 10.1016/j.antiviral.2015.03.015Chen YC, Sheng J, Trang P, Liu F. Potential application of the CRISPR/Cas9 system against herpesvirus infections. Viruses. 2018; 10(6): 291. doi: 10.3390/v10060291Chen YC, Sheng J, Trang P, Liu F. Potential application of the CRISPR/Cas9 system against herpesvirus infections. Viruses. 2018; 10(6): 291. doi: 10.3390/v10060291Itzhaki RF. Corroboration of a major role for herpes simplex virus type 1 in Alzheimer’s disease. Front Aging Neurosci. 2018; 10: 324. doi: 10.3389/fnagi.2018.00324Itzhaki RF. Corroboration of a major role for herpes simplex virus type 1 in Alzheimer’s disease. Front Aging Neurosci. 2018; 10: 324. doi: 10.3389/fnagi.2018.00324Cohen JI, Fauci AS, Varmus H, Nabel GJ. Epstein – Barr virus: An important vaccine target for cancer prevention. Sci Transl Med. 2011; 3(107): 107fs7. doi: 10.1126/scitranslmed.3002878Cohen JI, Fauci AS, Varmus H, Nabel GJ. Epstein – Barr virus: An important vaccine target for cancer prevention. Sci Transl Med. 2011; 3(107): 107fs7. doi: 10.1126/scitranslmed.3002878Gergen J, Coulon F, Creneguy A, Elain-Duret N, Gutierrez A, Pinkenburg O, et al. Multiplex CRISPR/Cas9 system impairs HCMV replication by excising an essential viral gene. PLoS One. 2018; 13(2): e0192602. doi: 10.1371/journal.pone.0192602Gergen J, Coulon F, Creneguy A, Elain-Duret N, Gutierrez A, Pinkenburg O, et al. Multiplex CRISPR/Cas9 system impairs HCMV replication by excising an essential viral gene. PLoS One. 2018; 13(2): e0192602. doi: 10.1371/journal.pone.0192602Gravitt PE. Evidence and impact of human papillomavirus latency. Open Virol J. 2012; 6: 198-203. doi: 10.2174/1874357901206010198Gravitt PE. Evidence and impact of human papillomavirus latency. Open Virol J. 2012; 6: 198-203. doi: 10.2174/1874357901 206010198Hu Z, Yu L, Zhu D, Ding W, Wang X, Zhang C, et al. Disruption of HPV16-E7 by CRISPR/Cas system induces apoptosis and growth inhibition in HPV16 positive human cervical cancer cells. Biomed Res Int. 2014: 612823. doi: 10.1155/2014/612823Hu Z, Yu L, Zhu D, Ding W, Wang X, Zhang C, et al. Disruption of HPV16-E7 by CRISPR/Cas system induces apoptosis and growth inhibition in HPV16 positive human cervical cancer cells. Biomed Res Int. 2014: 612823. doi: 10.1155/2014/612823

The authors declare that there are no conflicts of interest present.