Animal models for IL-5 function and for each of the suggested disease targets exist Cook et al. Ribozyme expression and activity levels in Stransduced clonal CEM cell lines. The plot of the fraction of uncleaved substrate versus time was fit to a double exponential curve using the KaleidaGraph graphing program Synergy Software, Reading, PA. However, mouse targeted ribozymes are useful to test efficacy of action of the ribozyme prior to testing in humans. The support was washed with 1. The above ribozyme transcription units can be incorporated into a variety of vectors for introduction into mammalian cells, including but not restricted to, plasmid DNA vectors, viral DNA vectors such as adenovirus or adeno-associated virus vectors , or viral RNA vectors such as retroviral, or alpha virus vectors. Each subunit bears a stretch of amino acids that is homologous to the oncogene, v-rel.

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Other sequences in the 5′ portion of the bcr mRNA or the 3′ portion of the abl mRNA may also be targeted for ribozyme cleavage. This timing of treatment will reduce the number of affected cells and disable cellular replication. In this invention small enzymatic nucleic acid motifs e.

Cellular lanes contain total cellular RNA from cells transfected with one of the four self-processing constructs. Current treatment options are limited to antibiotics to reduce the bacterial load and non-steroidal anti-inflammatories to reduce fever.

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The nucleotide used can be any normal nucleotide or may be substituted in various positions by methods well known in the art, e. If desired, the first ribozyme may be encoded on a separate vector and may have. An “aryl” group refers to an aromatic group which has at least one ring having a conjugated n electron system and includes carbocyclic aryl, heterocyclic aryl and biaryl groups, all of which may be optionally substituted.


Figure 72 is a graph showing RNA cleavage by hairpin ribozymes targeted to site M. Inhibition of IL-5 expression in the lungs can decrease the activation of eosinophils and will help alleviate the symptoms of asthma. Septic shock, with an incidence ofcases per year in the United States, is the major cause of death in intensive care units.

The term effective means that available amount of chemical actually provided to the reaction mixture that is able to react with the other reagents present in the mixture. This assay measures the level of expression from the type 2 pol promoter. The hairpin ribozyme is targeted against site J.

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The role of cytokines in the pathogenesis of psoriasis has been investigated. Lower case letters indicate vector sequence present in the RNA when transcribed from a Nindlll-linearized plasmid. Those in the art are familiar with such steps. Transfection of ribozymes into 53g.drover In a preferred embodiment, the conditions are dibutyltin oxide, tetrabutylammonium fluoride and SEM-CI. While primarily affecting children under two years of age.

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The ability of the ribozymes to inhibit TNF-a secretion and lethal shock caused by systemic LPS administration are assessed. Positions of 2′-hydroxyl group substitution is 15 indicated. Those of ordinary skill in the art, will find that it is clear from the examples described that other ribozymes that cleave these sites in RSV mRNAs encoding 1C, 1 B and Wcamm proteins may be readily designed and are within the invention.


For instance, the appropriate adenovirus vector can be administered by direct injection into the synovium: Each subunit bears a stretch of amino acids that is homologous to the oncogene, v-rel. By delivered is meant that the actual amount of chemical in the reaction mix is known. If probes of 53gg.driver specific activity are used for both transcripts, then a qualitative comparison of RNA levels will be adequate and will ‘ decrease the cost of the initial diagnosis.

CE is cyanoethyl, DMT is a standard blocking group. In step 1, cn base protecting groups and cyanoethyl groups are removed by standard procedure.

Ribozyme inhibition of ICAM-1 expression can reduce transplant rejection and alleviate symptoms in patients with rheumatoid arthritis, asthma or other acute and chronic inflammatory disorders. As shown in the figure 14, ribozymes that are deprotected using one pot protocol have 53g.drived activity comparable to ribozymes that are deprotected using a two pot protocol.

Wcqm correlation lends support to the hypothesis that reduced viral replication is physiologically linked to TNF-a levels: In general, the invention features a method for preparing multi-copy cassettes encoding a defined ribozyme structure for production of a ribozyme at a decreased cost. RNA isolation and cleavage reactions were as described in Fig. In this aspect, the process according to a second major embodiment, comprises: In addition, NF-xB is required for the expression of the oncogene c-myb F.