Further development of direct-acting antiviral agents against human SARS-CoV-2 infections remains a public health priority. Here, we report that an antisense peptide-conjugated morpholino oligomer (PPMO) named 5′END-2, targeting a highly conserved sequence in the 5′ UTR of SARS-CoV-2 genomic RNA, potently suppressed SARS-CoV-2 growth in vitro and in vivo. In HeLa-ACE 2 cells, 5′END-2 produced IC50 values of between 40 nM and 1.15 μM in challenges using six genetically disparate strains of SARS-CoV-2, including JN.1. In vivo, using K18-hACE2 mice and the WA-1/2020 virus isolate, two doses of 5′END-2 at 10 mg/kg, administered intranasally on the day before and the day after infection, produced approximately 1.4 log10 virus titer reduction in lung tissue at 3 days post-infection. Under a similar dosing schedule, intratracheal administration of 1.0–2.0 mg/kg 5′END-2 produced over 3.5 log10 virus growth suppression in mouse lungs. Electrophoretic mobility shift assays characterized specific binding of 5′END-2 to its complementary target RNA. Furthermore, using reporter constructs containing SARS-CoV-2 5′ UTR leader sequence, in an in-cell system, we observed that 5′END-2 could interfere with translation in a sequence-specific manner. The results demonstrate that direct pulmonary delivery of 5′END-2 PPMO is a promising antiviral strategy against SARS-CoV-2 infections and warrants further development.
", "doi": "10.1016/j.omtn.2024.102331", "issn": "2162-2531", "publisher": "Cell Press", "publication": "Moleclar Therapy - Nucleic Acids", "publication_date": "2024-12-10", "series_number": "4", "volume": "35", "issue": "4", "pages": "102331" }, { "id": "authors:q1s9m-d2498", "collection": "authors", "collection_id": "q1s9m-d2498", "cite_using_url": "https://authors.library.caltech.edu/records/q1s9m-d2498", "type": "article", "title": "Genome organization around nuclear speckles drives mRNA splicing efficiency", "author": [ { "family_name": "Bhat", "given_name": "Prashant", "orcid": "0000-0003-3832-4871", "clpid": "Bhat-Prashant" }, { "family_name": "Chow", "given_name": "Amy", "clpid": "Chow-Amy" }, { "family_name": "Emert", "given_name": "Benjamin", "orcid": "0000-0002-4857-2425", "clpid": "Emert-Benjamin" }, { "family_name": "Ettlin", "given_name": "Olivia", "orcid": "0000-0002-9243-9752", "clpid": "Ettlin-Olivia" }, { "family_name": "Quinodoz", "given_name": "Sofia A.", "orcid": "0000-0003-1862-5204", "clpid": "Quinodoz-Sofia-A" }, { "family_name": "Strehle", "given_name": "Mackenzie", "orcid": "0000-0003-1410-8701", "clpid": "Strehle-Mackenzie" }, { "family_name": "Takei", "given_name": "Yodai", "orcid": "0000-0002-7226-5185", "clpid": "Takei-Yodai" }, { "family_name": "Burr", "given_name": "Alex", "orcid": "0000-0003-2741-9592", "clpid": "Burr-Alex" }, { "family_name": "Goronzy", "given_name": "Isabel N.", "orcid": "0000-0002-6713-9192", "clpid": "Goronzy-Isabel-N" }, { "family_name": "Chen", "given_name": "Allen W.", "orcid": "0000-0002-6351-2578", "clpid": "Chen-Allen-W" }, { "family_name": "Huang", "given_name": "Wesley", "orcid": "0000-0002-6483-8297", "clpid": "Huang-Wesley" }, { "family_name": "Ferrer", "given_name": "Jose Lorenzo M.", "orcid": "0000-0002-3405-7596", "clpid": "Ferrer-Jose-Lorenzo-M" }, { "family_name": "Soehalim", "given_name": "Elizabeth", "clpid": "Soehalim-Elizabeth" }, { "family_name": "Goh", "given_name": "Say-Tar", "clpid": "Goh-Say-Tar" }, { "family_name": "Chari", "given_name": "Tara", "orcid": "0000-0002-6953-4313", "clpid": "Chari-Tara" }, { "family_name": "Sullivan", "given_name": "Delaney K.", "orcid": "0000-0002-8359-6705", "clpid": "Sullivan-Delaney-K" }, { "family_name": "Blanco", "given_name": "Mario R.", "orcid": "0000-0002-9852-2231", "clpid": "Blanco-Mario-R" }, { "family_name": "Guttman", "given_name": "Mitchell", "orcid": "0000-0003-4748-9352", "clpid": "Guttman-M" } ], "abstract": "The nucleus is highly organized, such that factors involved in the transcription and processing of distinct classes of RNA are confined within specific nuclear bodies1,2. One example is the nuclear speckle, which is defined by high concentrations of protein and noncoding RNA regulators of pre-mRNA splicing3. What functional role, if any, speckles might play in the process of mRNA splicing is unclear4,5. Here we show that genes localized near nuclear speckles display higher spliceosome concentrations, increased spliceosome binding to their pre-mRNAs and higher co-transcriptional splicing levels than genes that are located farther from nuclear speckles. Gene organization around nuclear speckles is dynamic between cell types, and changes in speckle proximity lead to differences in splicing efficiency. Finally, directed recruitment of a pre-mRNA to nuclear speckles is sufficient to increase mRNA splicing levels. Together, our results integrate the long-standing observations of nuclear speckles with the biochemistry of mRNA splicing and demonstrate a crucial role for dynamic three-dimensional spatial organization of genomic DNA in driving spliceosome concentrations and controlling the efficiency of mRNA splicing.
\n