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At least 91 records · Page 5Linked to original sources
PubMed1998

Antisense RNA complementary to hepatitis B virus specifically inhibits viral replication.

BACKGROUND & AIMS: Chronic infection with the hepatitis B virus (HBV) is a major public health problem, and currently available therapies have limited efficacy. Gene therapy strategies for HBV infection are under active investigation. We evaluated the potential of antisense RNA transcribed from antisense genes to interfere with HBV replication. METHODS: Subgenomic fragments of the HBV genome were studied with respect to the property of inhibiting HBV replication when intracellularly expressed in the antisense orientation. RESULTS: Antisense RNAs derived from the HBV genome specifically inhibited HBV replication and antigen expression in human hepatocellular carcinoma cells by 60%-75%. DNA sequences corresponding to the identified RNAs had no effect on HBV replication, indicating that inhibitory effects are mediated by RNA. Transcripts corresponding to the inhibitory subgenomic fragments were present at high levels. One antisense RNA was found to reduce the amount of pregenomic RNA encapsidated into core particles as a molecular mechanism of antiviral effects. CONCLUSIONS: Certain antisense RNA molecules will have substantial antiviral effects against HBV. Antisense RNAs derived from the HBV genome are promising candidates as antiviral agents and may serve as novel tools to identify functionally important regions of HBV transcripts.

Base Sequence↗
PubMed1981

Detection of RNA complementary to herpes simplex virus DNA in human cervical squamous cell neoplasms.

Nonneoplastic and neoplastic cervical biopsy specimens were examined by in situ hybridization to 125I-labeled DNA of herpes simplex virus (HSV), adenovirus, and bacteriophage lambda DNA's, and quantitative hybridization data were obtained using a Video Image Analyser. HSV-specific RNA was detected in 72% of cervical intraepithelial neoplasia, 60% of squamous cervical carcinomas, 2% of nonneoplastic cervices, and 9% of primary adenocarcinomas of the cervix. None of the tissues gave positive hybridization with adenovirus or lambda DNA probes. In paired biopsies of cervical intraepithelial neoplasia and nonneoplastic epithelium from 29 individuals, HSV-specific RNA was detected only in the epithelium of the neoplastic sample and not in the nonneoplastic control. Infectious HSV-2 was isolated from a low proportion (2%) of both ectocervical swabs and cell-free tissue extracts of patients examined, suggesting that the HSV-specific RNA detected in squamous cell neoplasms was not due to overt infections.

Antibodies, Viral↗
PubMed1990

Coding strategy of rice stripe virus: major nonstructural protein is encoded in viral RNA segment 4 and coat protein in RNA complementary to segment 3.

The two major proteins found in plants infected with rice stripe virus (RSV), coat protein and a major nonstructural protein (major NS), were purified and their partial amino acid sequences were determined. Oligonucleotides were synthesized according to the amino acid sequence information, and used as probes for Northern blot analyses of four single-stranded RNA species (segments 1-4) and four double-stranded RNA species of RSV. The results indicated that the coding strategy of RSV was similar to those of ambisense viruses: Segment 3 coded for the coat protein in the (-) sense while segment 4 coded for the major NS protein in the (+) sense.

Amino Acid Sequence↗
PubMed1991

Inhibition of Moloney murine leukemia virus-induced leukemia in transgenic mice expressing antisense RNA complementary to the retroviral packaging sequences.

Recombinant plasmids pLP psi as and pCP psi as were constructed by positioning the Moloney murine leukemia virus (M-MuLV) proviral packaging (psi) sequences in reverse orientation under the transcriptional regulation of lymphotropic promoter/regulatory elements from the M-MuLV long terminal repeat or the cytomegalovirus immediate-early region. Linear fragments containing the antisense psi and the appropriate transcriptional regulatory sequences from these plasmids were introduced into the mouse germ line by zygote microinjection. The chromosomal integration, germ-line transmission, and lymphocyte-directed expression of the antisense psi RNA were confirmed. Control (nontransgenic) and transgenic mice containing either the pLP psi as or the pCP psi as sequences were infected with M-MuLV on the day of birth and assayed for signs of leukemia between 12 and 14 weeks of age with standard assay procedures. While 31% (11 of 36) of the control, nontransgenic, mice developed leukemia, none of the antisense psi transgenic mice developed any symptoms of leukemia. The pCP psi as sequences were also introduced into mouse NIH 3T3 cells and stably transformed cell lines were isolated. When infected with M-MuLV these cells were shown to produce virus devoid of packaged viral RNA.

Animals↗
PubMed1987

Transcriptional analysis of the cyanobacterial gvpABC operon in differentiated cells: occurrence of an antisense RNA complementary to three overlapping transcripts.

Cyanobacteria are photosynthetic prokaryotes able to colonize almost all kinds of ecosystems. Some of them exhibit differentiation processes and/or may establish tight symbiotic associations. Upon changes in the environmental conditions, the cyanobacterium Calothrix 7601 differentiates hormogonia which are short filaments of small cells resulting from cellular division and fragmentation of the long filaments of vegetative cells. In Calothrix, hormogonia are characterized by their gliding motility and by a massive production of gas vesicles which confer buoyancy. At least four genes are involved in the formation of gas vesicles, three of which are organized in one operon (gvpABC). Four different RNA species, only present after induction of hormogonia differentiation, result from transcription of this operon. Mapping of the 5' and 3' ends of these transcripts demonstrates the presence of gvpA, gvpAB and gvpABC transcripts, all three having the same 5' end. Each of the three transcripts terminates a few bases downstream from stem-and-loop structures. Most interestingly, the fourth transcript is an antisense RNA starting from the 3' end of the gvpB gene and ending within the gvpA gene. This antisense RNA can thus form an homologous duplex with the three other transcripts, thereby being able to impair translation and/or modify mRNA stability.

Chromosome Mapping↗
PubMed1990

Maternal mRNA from clam oocytes can be specifically unmasked in vitro by antisense RNA complementary to the 3'-untranslated region.

Clam oocytes display a striking difference in the pattern of protein synthesis on fertilization, which is maintained when cell-free extracts are assayed in the rabbit reticulocyte lysate. The mRNAs encoding ribonucleotide reductase and cyclin A (the major masked messages in the oocyte extracts) can be translationally activated by gel filtration in 0.5 M KCl, presumably by removal of repressor protein(s). When synthetic RNAs corresponding to different segments of ribonucleotide reductase and cyclin A mRNAs were added to oocyte extracts in 0.5 M KCl in a "competitive unmasking" assay, specific and complete unmasking of these mRNAs occurred on addition of antisense 3'-noncoding transcripts. The unmasking region in ribonucleotide reductase mRNA maps to a region of 134 nucleotides centered one-third of the way down the 3'-noncoding region.

Animals↗
PubMed1983

Oligomeric forms of potato spindle tuber viroid (PSTV) and of its complementary RNA are present in nuclei isolated from viroid-infected potato cells.

Different oligomeric forms of PSTV are detected in nuclei isolated from PSTV-infected potato cells by means of molecular hybridization, using as probes synthetic oligodeoxyribonucleotides with sequence specificity for (+)PSTV and for (-)PSTV. In addition to several species of longer-than-unit-length (-)PSTV molecules, two oligomeric forms of (+)PSTV are detected, which correspond in size to RNA strands of approximately two and three times viroid unit-length. They must be considered as the precursors of the circular and linear (+)PSTV monomers accumulating in the cell nucleus.

Base Sequence↗