{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:44:56Z","timestamp":1760060696307,"version":"build-2065373602"},"reference-count":90,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2025,9,12]],"date-time":"2025-09-12T00:00:00Z","timestamp":1757635200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"University of California-Berkeley"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Nucleic acid-based gene-interfering molecules, such as antisense oligonucleotides, ribozymes, and small interfering RNA (siRNA), represent exciting gene-targeting agents for therapeutic applications. RNase P ribozymes derived from M1 RNA, the catalytic RNA subunit of RNase P in Escherichia coli, have shown great promise as a novel nucleic acid-based gene interference approach to modulate gene expression. When M1 RNA is covalently linked to a guide sequence (GS), it can be engineered into a sequence-specific endonuclease M1GS ribozyme, which can hydrolyze any mRNA that base-pairs with the guide sequence. M1GS activity enhancement has been achieved through an in vitro selection process that introduced mutations into M1 RNA. This selection process generated ribozyme variants with improved cleavage efficiency and substrate affinity. Hepatitis B virus (HBV) chronically infects more than 250 million people worldwide and is the leading cause of cirrhosis and liver cancer globally. Current FDA-approved drugs cannot completely eliminate HBV chronic infections. RNase P ribozymes have recently been demonstrated to effectively inhibit HBV gene expression and replication in human cells. This review summarizes the recent progress in using RNase P ribozymes to inhibit HBV infection and discusses prospects for developing engineered RNase P ribozymes for therapeutic applications against HBV infection and associated diseases.<\/jats:p>","DOI":"10.3390\/molecules30183725","type":"journal-article","created":{"date-parts":[[2025,9,15]],"date-time":"2025-09-15T10:51:33Z","timestamp":1757933493000},"page":"3725","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Potential Applications of RNase P Ribozyme Against Hepatitis B Virus"],"prefix":"10.3390","volume":"30","author":[{"given":"Thomas","family":"Sorrell","sequence":"first","affiliation":[{"name":"School of Public Health, University of California, Berkeley, CA 94720, USA"},{"name":"Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yujun","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Public Health, University of California, Berkeley, CA 94720, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fenyong","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Public Health, University of California, Berkeley, CA 94720, USA"},{"name":"Program in Comparative Biochemistry, University of California, Berkeley, CA 94720, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1016\/S2468-1253(18)30056-6","article-title":"Global prevalence, treatment, and prevention of hepatitis B virus infection in 2016: A modelling study","volume":"3","author":"Gamkrelidze","year":"2018","journal-title":"Lancet Gastroenterol. Hepatol."},{"key":"ref_2","first-page":"641","article-title":"Hepadnaviridae","volume":"Volume 1","author":"Knipe","year":"2021","journal-title":"Fields Virology: DNA Viruses"},{"key":"ref_3","unstructured":"World Health Organization (2025, June 01). Hepatitis B. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/hepatitis-b."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"332","DOI":"10.1016\/S2468-1253(22)00386-7","article-title":"Global reporting of progress towards elimination of hepatitis B and hepatitis C","volume":"8","author":"Cui","year":"2023","journal-title":"Lancet Gastroenterol. Hepatol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1030","DOI":"10.4254\/wjh.v7.i8.1030","article-title":"Current and future antiviral drug therapies of hepatitis B chronic infection","volume":"7","author":"Koumbi","year":"2015","journal-title":"World J. Hepatol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1457","DOI":"10.1038\/nbt915","article-title":"Approaches for the sequence-specific knockdown of mRNA","volume":"21","author":"Scherer","year":"2003","journal-title":"Nat. Biotechnol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e114760","DOI":"10.15252\/embj.2023114760","article-title":"RNA-based medicine: From molecular mechanisms to therapy","volume":"42","author":"Sparmann","year":"2023","journal-title":"EMBO J."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"eadd8643","DOI":"10.1126\/science.add8643","article-title":"CRISPR technology: A decade of genome editing is only the beginning","volume":"379","author":"Wang","year":"2023","journal-title":"Science"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1016\/j.bbaexp.2007.09.001","article-title":"Inhibition of gene expression in human cells using RNase P-derived ribozymes and external guide sequences","volume":"1769","author":"Kim","year":"2007","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"6759","DOI":"10.1074\/jbc.R100067200","article-title":"RNase P: Variations and uses","volume":"277","author":"Gopalan","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Kirsebom, L.A., Liu, F., and McClain, W.H. (2024). The discovery of a catalytic RNA within RNase P and its legacy. J. Biol. Chem., 300.","DOI":"10.1016\/j.jbc.2024.107318"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Campos-Valdez, M., Monroy-Ram\u00edrez, H.C., Armend\u00e1riz-Borunda, J., and S\u00e1nchez-Orozco, L.V. (2021). Molecular Mechanisms during Hepatitis B Infection and the Effects of the Virus Variability. Viruses, 13.","DOI":"10.3390\/v13061167"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Brandes, N., and Linial, M. (2016). Gene overlapping and size constraints in the viral world. Biol. Direct, 11.","DOI":"10.1186\/s13062-016-0128-3"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1053\/j.gastro.2018.07.058","article-title":"Insights from deep sequencing of the HBV genome\u2014Unique, tiny, and misunderstood","volume":"156","author":"McNaughton","year":"2019","journal-title":"Gastroenterology"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Li, S., Wang, Z., Li, Y., and Ding, G. (2017). Adaptive evolution of proteins in hepatitis B virus during divergence of genotypes. Sci. Rep., 7.","DOI":"10.1038\/s41598-017-02012-8"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1006\/jmbi.2000.4481","article-title":"Structural organization of the hepatitis B virus minichromosome","volume":"307","author":"Bock","year":"2001","journal-title":"J. Mol. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Boonstra, A., and Sari, G. (2025). HBV cccDNA: The Molecular Reservoir of Hepatitis B Persistence and Challenges to Achieve Viral Eradication. Biomolecules, 15.","DOI":"10.3390\/biom15010062"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Tu, T., Budzinska, M.A., Shackel, N.A., and Urban, S. (2017). HBV DNA Integration: Molecular Mechanisms and Clinical Implications. Viruses, 9.","DOI":"10.3390\/v9040075"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1007\/s12072-017-9829-7","article-title":"Hepatitis B virus: Virology, molecular biology, life cycle and intrahepatic spread","volume":"11","author":"Karayiannis","year":"2017","journal-title":"Hepatol. Int."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/j.antiviral.2015.09.011","article-title":"Hepatitis B virus reverse transcriptase\u2014Target of current antiviral therapy and future drug development","volume":"123","author":"Clark","year":"2015","journal-title":"Antivir. Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.coviro.2014.04.005","article-title":"Nucleoside\/nucleotide analog inhibitors of hepatitis B virus polymerase: Mechanism of action and resistance","volume":"8","author":"Pacheco","year":"2014","journal-title":"Curr. Opin. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1221","DOI":"10.1126\/science.1243462","article-title":"Specific and nonhepatotoxic degradation of nuclear hepatitis B virus cccDNA","volume":"343","author":"Lucifora","year":"2014","journal-title":"Science"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1038\/nsmb.1744","article-title":"APOBEC3 proteins mediate the clearance of foreign DNA from human cells","volume":"17","author":"Stenglein","year":"2010","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Inoue, J., Sato, K., Ninomiya, M., and Masamune, A. (2021). Envelope Proteins of Hepatitis B Virus: Molecular Biology and Involvement in Carcinogenesis. Viruses, 13.","DOI":"10.3390\/v13061124"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.addr.2007.03.005","article-title":"RNAi therapeutics: Principles, prospects and challenges","volume":"59","author":"Aagaard","year":"2007","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"17589","DOI":"10.3390\/ijms160817589","article-title":"Progress and Prospects of Anti-HBV Gene Therapy Development","volume":"16","author":"Maepa","year":"2015","journal-title":"Int. J. Mol. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"973","DOI":"10.1038\/mt.2013.31","article-title":"Hepatocyte-targeted RNAi therapeutics for the treatment of chronic hepatitis B virus infection","volume":"21","author":"Wooddell","year":"2013","journal-title":"Mol. Ther."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Caffrey, D.R., Zhao, J., Song, Z., Schaffer, M.E., Haney, S.A., Subramanian, R.R., Seymour, A.B., and Hughes, J.D. (2011). siRNA off-target effects can be reduced at concentrations that match their individual potency. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0021503"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1892","DOI":"10.1073\/pnas.0308698100","article-title":"Short interfering RNAs can induce unexpected and divergent changes in the levels of untargeted proteins in mammalian cells","volume":"101","author":"Scacheri","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1779","DOI":"10.1007\/s00018-019-03021-8","article-title":"Orthologous CRISPR\/Cas9 systems for specific and efficient degradation of covalently closed circular DNA of hepatitis B virus","volume":"76","author":"Kostyushev","year":"2019","journal-title":"Cell Mol. Life Sci."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Martinez, M.G., Smekalova, E., Combe, E., Gregoire, F., Zoulim, F., and Testoni, B. (2022). Gene Editing Technologies to Target HBV cccDNA. Viruses, 14.","DOI":"10.3390\/v14122654"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"91","DOI":"10.3748\/wjg.v8.i1.91","article-title":"Anti-HBV hairpin ribozyme-mediated cleavage of target RNA in vitro","volume":"8","author":"Song","year":"2002","journal-title":"World J. Gastroenterol."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Smith, A., Zhang, I., Trang, P., and Liu, F. (2024). Engineering of RNase P Ribozymes for Therapy against Human Cytomegalovirus Infection. Viruses, 16.","DOI":"10.3390\/v16081196"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2909","DOI":"10.1016\/j.ymthe.2022.05.013","article-title":"Targeting the hepatitis B cccDNA with a sequence-specific ARCUS nuclease to eliminate hepatitis B virus","volume":"30","author":"Gorsuch","year":"2022","journal-title":"Mol. Ther."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Weber, N.D., Stone, D., Sedlak, R.H., De Silva Feelixge, H.S., Roychoudhury, P., Schiffer, J.T., Aubert, M., and Jerome, K.R. (2014). AAV-mediated delivery of zinc finger nucleases targeting hepatitis B virus inhibits active replication. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0097579"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"947","DOI":"10.1038\/mt.2010.20","article-title":"Zinc-finger Nucleases as a Novel Therapeutic Strategy for Targeting Hepatitis B Virus DNAs","volume":"18","author":"Cradick","year":"2010","journal-title":"Mol. Ther."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1038\/mt.2013.212","article-title":"An efficient antiviral strategy for targeting hepatitis B virus genome using transcription activator-like effector nucleases","volume":"22","author":"Chen","year":"2014","journal-title":"Mol. Ther."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1725","DOI":"10.1038\/s41591-021-01513-4","article-title":"Safety, tolerability and antiviral activity of the antisense oligonucleotide bepirovirsen in patients with chronic hepatitis B: A phase 2 randomized controlled trial","volume":"27","author":"Yuen","year":"2021","journal-title":"Nat. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1957","DOI":"10.1056\/NEJMoa2210027","article-title":"Efficacy and Safety of Bepirovirsen in Chronic Hepatitis B Infection","volume":"387","author":"Yuen","year":"2022","journal-title":"N. Engl. J. Med."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"404","DOI":"10.1016\/j.jhep.2024.03.046","article-title":"JNJ-73763989 and bersacapavir treatment in nucleos(t)ide analogue-suppressed patients with chronic hepatitis B: REEF-2","volume":"81","author":"Agarwal","year":"2024","journal-title":"J. Hepatol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1007\/s40259-024-00694-x","article-title":"Investigational RNA Interference Agents for Hepatitis B","volume":"39","author":"Hui","year":"2025","journal-title":"BioDrugs"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1016\/j.omtn.2020.03.005","article-title":"Permanent Inactivation of HBV Genomes by CRISPR\/Cas9-Mediated Non-cleavage Base Editing","volume":"20","author":"Yang","year":"2020","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Yan, B., Liu, Y., Chen, Y.C., and Liu, F. (2023). A RNase P Ribozyme Inhibits Gene Expression and Replication of Hepatitis B Virus in Cultured Cells. Microorganisms, 11.","DOI":"10.3390\/microorganisms11030654"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1146\/annurev.biochem.67.1.153","article-title":"Ribonuclease P: Unity and diversity in a tRNA processing ribozyme","volume":"67","author":"Frank","year":"1998","journal-title":"Annu. Rev. Biochem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1146\/annurev.biochem.71.110601.135352","article-title":"Eukaryotic ribonuclease P: A plurality of ribonucleoprotein enzymes","volume":"71","author":"Xiao","year":"2002","journal-title":"Annu. Rev. Biochem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1261\/rna.079475.122","article-title":"Human RNase P: Overview of a ribonuclease of interrelated molecular networks and gene-targeting systems","volume":"29","author":"Jarrous","year":"2023","journal-title":"RNA"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"9409","DOI":"10.1021\/bi980613c","article-title":"The protein component of Bacillus subtilis ribonuclease P increases catalytic efficiency by enhancing interactions with the 5\u2032 leader sequence of pre-tRNAAsp","volume":"37","author":"Crary","year":"1998","journal-title":"Biochemistry"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"818","DOI":"10.1006\/jmbi.1997.0906","article-title":"Analysis of the functional role of conserved residues in the protein subunit of ribonuclease P from Escherichia coli","volume":"267","author":"Gopalan","year":"1997","journal-title":"J. Mol. Biol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1016\/0092-8674(83)90117-4","article-title":"The RNA moiety of ribonuclease P is the catalytic subunit of the enzyme","volume":"35","author":"Gardiner","year":"1983","journal-title":"Cell"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1002\/bip.10521","article-title":"Roles of protein subunits in RNA-protein complexes: Lessons from ribonuclease P","volume":"73","author":"Hsieh","year":"2004","journal-title":"Biopolymers"},{"key":"ref_51","first-page":"613","article-title":"The protein cofactor allows the sequence of an RNase P ribozyme to diversify by maintaining the catalytically active structure of the enzyme","volume":"3","author":"Kim","year":"1997","journal-title":"RNA"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"9545","DOI":"10.1021\/bi025553w","article-title":"The affinity of magnesium binding sites in the Bacillus subtilis RNase P x pre-tRNA complex is enhanced by the protein subunit","volume":"41","author":"Kurz","year":"2002","journal-title":"Biochemistry"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"2393","DOI":"10.1021\/bi972530m","article-title":"Protein component of Bacillus subtilis RNase P specifically enhances the affinity for precursor-tRNAAsp","volume":"37","author":"Kurz","year":"1998","journal-title":"Biochemistry"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"15212","DOI":"10.1073\/pnas.95.26.15212","article-title":"Protein component of the ribozyme ribonuclease P alters substrate recognition by directly contacting precursor tRNA","volume":"95","author":"Niranjanakumari","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1006\/jmbi.1996.0272","article-title":"Mapping in three dimensions of regions in a catalytic RNA protected from attack by an Fe(II)-EDTA reagent","volume":"258","author":"Westhof","year":"1996","journal-title":"J. Mol. Biol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"784","DOI":"10.1038\/nature09516","article-title":"Structure of a bacterial ribonuclease P holoenzyme in complex with tRNA","volume":"468","author":"Reiter","year":"2010","journal-title":"Nature"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"5120","DOI":"10.1038\/s41467-022-32843-7","article-title":"Structural and mechanistic basis for recognition of alternative tRNA precursor substrates by bacterial ribonuclease P","volume":"13","author":"Zhu","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1017\/S1355838201010469","article-title":"Function and subnuclear distribution of Rpp21, a protein subunit of the human ribonucleoprotein ribonuclease P","volume":"7","author":"Jarrous","year":"2001","journal-title":"RNA"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1126\/science.1697102","article-title":"External guide sequences for an RNA enzyme","volume":"249","author":"Forster","year":"1990","journal-title":"Science"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1126\/science.2443980","article-title":"Model substrates for an RNA enzyme","volume":"238","author":"McClain","year":"1987","journal-title":"Science"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"8006","DOI":"10.1073\/pnas.89.17.8006","article-title":"Targeted cleavage of mRNA by human RNase P","volume":"89","author":"Yuan","year":"1992","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"995","DOI":"10.1038\/mt.2013.37","article-title":"Inhibition of Hepatitis B Virus Gene Expression and Replication by Ribonuclease P","volume":"21","author":"Xia","year":"2013","journal-title":"Mol. Ther."},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Zhang, Z., Vu, G.P., Gong, H., Xia, C., Chen, Y.C., Liu, F., Wu, J., and Lu, S. (2013). Engineered external guide sequences are highly effective in inhibiting gene expression and replication of hepatitis B virus in cultured cells. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0065268"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"3185","DOI":"10.1073\/pnas.89.8.3185","article-title":"Targeted cleavage of mRNA in vitro by RNase P from Escherichia coli","volume":"89","author":"Li","year":"1992","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1101\/gad.9.4.471","article-title":"Inhibition of viral gene expression by the catalytic RNA subunit of RNase P from Escherichia coli","volume":"9","author":"Liu","year":"1995","journal-title":"Genes Dev."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1016\/S1097-2765(03)00357-5","article-title":"Eukaryotic RNase P: Role of RNA and Protein Subunits of a Primordial Catalytic Ribonucleoprotein in RNA-Based Catalysis","volume":"12","author":"Mann","year":"2003","journal-title":"Mol. Cell"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"5120","DOI":"10.1093\/nar\/gki828","article-title":"RNase P: Role of distinct protein cofactors in tRNA substrate recognition and RNA-based catalysis","volume":"33","author":"Sharin","year":"2005","journal-title":"Nucleic Acids Res."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1038\/scientificamerican1292-90","article-title":"Directed molecular evolution","volume":"267","author":"Joyce","year":"1992","journal-title":"Sci. Am."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/0968-0004(92)90242-2","article-title":"In vitro genetics","volume":"17","author":"Szostak","year":"1992","journal-title":"Trends Biochem. Sci."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"763","DOI":"10.1146\/annurev.bi.64.070195.003555","article-title":"Diversity of oligonucleotide functions","volume":"64","author":"Gold","year":"1995","journal-title":"Annu. Rev. Biochem."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"10611","DOI":"10.1074\/jbc.275.14.10611","article-title":"RNase P ribozymes selected in vitro to cleave a viral mRNA effectively inhibit its expression in cell culture","volume":"275","author":"Kilani","year":"2000","journal-title":"J. Biol. Chem."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1007\/978-1-0716-3918-4_26","article-title":"In Vitro Amplification and Selection of Engineered RNase P Ribozyme for Gene Targeting Applications","volume":"2822","author":"Trang","year":"2024","journal-title":"Methods Mol. Biol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"32063","DOI":"10.1074\/jbc.M403059200","article-title":"Engineered RNase P ribozymes increase their cleavage activities and efficacies in inhibiting viral gene expression in cells by enhancing the rate of cleavage and binding of the target mRNA","volume":"279","author":"Zou","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"1004","DOI":"10.1126\/science.8351515","article-title":"Antisense oligonucleotides as therapeutic agents\u2014Is the bullet really magical?","volume":"261","author":"Stein","year":"1993","journal-title":"Science"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1182\/blood.V95.3.731.003k28_731_737","article-title":"In vivo inhibition by a site-specific catalytic RNA subunit of RNase P designed against the BCR-ABL oncogenic products: A novel approach for cancer treatment","volume":"95","author":"Cobaleda","year":"2000","journal-title":"Blood"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"1222","DOI":"10.1126\/science.2107573","article-title":"Ribozymes as Potential Anti-HIV-1 Therapeutic Agents","volume":"247","author":"Sarver","year":"1990","journal-title":"Science"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"6340","DOI":"10.1073\/pnas.90.13.6340","article-title":"A hairpin ribozyme inhibits expression of diverse strains of human immunodeficiency virus type 1","volume":"90","author":"Yu","year":"1993","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1146\/annurev.biochem.67.1.99","article-title":"Modified oligonucleotides: Synthesis and strategy for users","volume":"67","author":"Verma","year":"1998","journal-title":"Annu. Rev. Biochem."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.chembiol.2011.12.008","article-title":"RNA-based therapeutics: Current progress and future prospects","volume":"19","author":"Burnett","year":"2012","journal-title":"Chem. Biol."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"3222","DOI":"10.1073\/pnas.1014975108","article-title":"Oral delivery of RNase P ribozymes by Salmonella inhibits viral infection in mice","volume":"108","author":"Bai","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"7269","DOI":"10.1073\/pnas.0912813107","article-title":"Salmonella-mediated delivery of RNase P-based ribozymes for inhibition of viral gene expression and replication in human cells","volume":"107","author":"Bai","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"13070","DOI":"10.1073\/pnas.1201620109","article-title":"Effective inhibition of cytomegalovirus infection by external guide sequences in mice","volume":"109","author":"Jiang","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"5071","DOI":"10.1093\/nar\/29.24.5071","article-title":"Effective inhibition of herpes simplex virus 1 gene expression and growth by engineered RNase P ribozyme","volume":"29","author":"Trang","year":"2001","journal-title":"Nucleic Acids Res."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"1494","DOI":"10.1002\/jbm.a.37705","article-title":"Influence of ionizable lipid tail length on lipid nanoparticle delivery of mRNA of varying length","volume":"112","author":"Mrksich","year":"2024","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"4799","DOI":"10.1038\/s41467-025-59489-5","article-title":"Fusogenic lipid nanoparticles for rapid delivery of large therapeutic molecules to exosomes","volume":"16","author":"Son","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1038\/s41573-024-00977-6","article-title":"The 60-year evolution of lipid nanoparticles for nucleic acid delivery","volume":"23","author":"Cullis","year":"2024","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"5257","DOI":"10.1021\/acs.molpharmaceut.5c00670","article-title":"Recent Developments of RNA Vaccines and Therapeutics: Reagents, Formulations, and Characterization","volume":"22","author":"Tomeh","year":"2025","journal-title":"Mol. Pharm."},{"key":"ref_88","doi-asserted-by":"crossref","unstructured":"Li, D., Hamadalnil, Y., and Tu, T. (2024). Hepatitis B Viral Protein HBx: Roles in Viral Replication and Hepatocarcinogenesis. Viruses, 16.","DOI":"10.3390\/v16091361"},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"2230","DOI":"10.1073\/pnas.91.6.2230","article-title":"Hepatitis B virus X protein inhibits p53 sequence-specific DNA binding, transcriptional activity, and association with transcription factor ERCC3","volume":"91","author":"Wang","year":"1994","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"996","DOI":"10.1016\/j.jhep.2011.02.015","article-title":"Hepatitis B virus X protein is essential to initiate and maintain virus replication after infection","volume":"55","author":"Lucifora","year":"2011","journal-title":"J. Hepatol."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/18\/3725\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:44:54Z","timestamp":1760035494000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/18\/3725"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,12]]},"references-count":90,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["molecules30183725"],"URL":"https:\/\/doi.org\/10.3390\/molecules30183725","relation":{},"ISSN":["1420-3049"],"issn-type":[{"type":"electronic","value":"1420-3049"}],"subject":[],"published":{"date-parts":[[2025,9,12]]}}}