Antisense mRNA

Mautino MR, Morgan RA (2000) Potent inhibition of human immunodeficiency virus type 1 reph-cation by conditionally replicating human immunodeficiency virus-based lentiviral vectors expressing envelope antisense mRNA, Hum Gene Ther 11 2025-2037... 

An alternative system, which effectively results in the introduction of antisense oligonucleotides into the cell, entails application of gene therapy. In this case, a gene, which when transcribed yields (antisense) mRNA of appropriate nucleotide sequence, is introduced into the cell by a retroviral or other appropriate vector. This approach, as applied to the treatment of cancer and AIDS, is being appraised in a number of trials. 

Translation arrest or nuclease digestion by exogenously applied antisense oligonucleotides or by antisense mRNA produced from DNA delivered by a plasmid. Heterogeneous nuclear RNA is hnRNA. 

Kimelman, D. and Kirschner, M. W., An antisense mRNA directs the covalent modification of the transcript encoding fibroblast growth factor in Xenopus oocytes, Cell, 59, 687, 1989. 

Grothe, C. and Meisinger, C., Fibroblast growth factor (FGF)-2 sense and antisense mRNA and FGF receptor type 1 mRNA are present in the embryonic and adult rat nervous system specific detection by nuclease protection assay, Neurosci. Lett., 197, 175, 1995. 

Borja, A.Z., Meijers, C. and Zeller, R. (1993) Expression of alternatively spliced bFGF first coding exons and antisense mRNAs during chicken embryogenesis. Dev. Biol. 157 110-118. 

A typical antisense mRNA would be 17-25 nucleotides long and would destroy the target mRNA by binding to it and invoking ribonuclease H activity. The synthetic RNAs are, however, also quickly destroyed by other ribonucle-... 

Figure 8.4.3 Sequence- and strain-specific transcription of DNA into an mRNA sequences. Single-strand sense mRNA is formed. Antisense mRNA could form a double strand with such a molecule.

Chen DQ, KoUi BK, Yadava N et al. Episom expression of lecific sense and antisense mRNAs in Leishmania amazonensis Modulation of gp63 level in promast tes and thdr infection of macrophages in vitro. Infect Immun 2000 68(l) 80-6. 

Synthesis of Sense and Antisense mRNA from the Recombinant Lambda... 

Recombinant lambda DNA isolated from the Xgt22 cDNA library described above is an efficient template for the synthesis of sense and antisense mRNA vitro by RNA polymerase T7 or SP6 respectively. Using a procedure by Melton et al. (21) and Nielsen et al. (22), 1-2 yg "of capped mRNA can be synthesized from each microgram of template DNA. 

Figure 4. Autoradiogram of [-methionine labeled peptides synthesized in rabbit reticulocyte lysate and immuno-precipitated by a rabbit anti-insulin serum. The RNAs used were 1 pg of Brome Mosaic Virus as a control (lane 1), no RNA (lane 2), 0.5 yg of polyA+ RNA isolated from rat pancreas (lane 3), 0.2 yg of mRNA synthesized by T7 polymerase (lane 4), 0.2 yg of polyA+ RNA combined with 1 yg of antisense mRNA synthesized by SP6 polymerase (lane 5), and 0.2 yg of synthetic mRNA combined with 1 yg of antisense mRNA. The RNAs used in lane 5 and 6 were heated at 70°C for 15 min and cooled slowly before the translation reaction.

Thirty-seven antisense transgenic lines of tobacco were produced. The antisense mRNA was made in those plants, but none of them showed substantial decrease in PLD activity. Sequence differences between the castor and tobacco PLD genes might be one of the possible explanations for the inefficient antisense suppression. 

The progeny from transgenic plants with lowered tomato CRYl mRNA and protein levels, obtained by using an antisense mRNA approach, show elongated hypocotyl and decreased anthocyanins accumulation under blue Hght, providing the first direct evidence of a cryptochrome role in tomato photomor-... 

---