By K. I. Berns, C. Giraud (auth.), Professor Kenneth I. Berns M.D., Ph.D., Catherine Giraud Ph.D. (eds.)
Human gene treatment holds nice promise for the treatment of many genetic ailments. which will in attaining this sort of remedy there are necessities. First, the affected gene needs to be cloned, its se quence decided and its law safely characterised. moment, an appropriate vector for the supply of an outstanding replica of the affected gene has to be to be had. For a vector to be of use numerous attributes are hugely fascinating: those comprise skill to hold the intact gene (although this can be both the genomic or the cDNA shape) in a solid shape, skill to introduce the gene into the specified telephone sort, skill to specific the brought gene in an effectively regulated demeanour for a longer time period, and a scarcity of toxicity for the recipient. additionally of shock is the frequency of telephone transformation and, in certain cases, the power to introduce the gene into nondividing stem cells. Sev eral animal viruses were demonstrated as power vectors, yet none has confirmed to have the entire wanted houses defined above. for instance, retroviruses are tough to propagate in enough titers, don't combine into nondividing cells, and are of outrage due to their oncogenic houses in a few hosts and since they combine at many websites within the genome and, therefore, are almost certainly insertional mutagens. also, genes brought through retroviral vectors are often expressed for really brief classes of time. A moment virus used as a vector in version structures has been adenovirus (Ad).
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Extra resources for Adeno-Associated Virus (AAV) Vectors in Gene Therapy
1984; LABow and BERNS 1988) and in vitro (CHIORINI et al. 1994a; HONG et al. 1992; NI et al. 1994). Furthermore, Rep68 and Rep78 have been shown to nick the duplex ITR in a site- and strand-specific manner. This The Roles of AAV Rep Proteins in Gene Expression and Targeted Integration 27 cleavage, which occurs at the terminal resolution site (TRS). permits the replication of the hairpin structure in a process referred to as terminal resolution (1M and MUZVCZKA 1990). In addition to their role in AAV DNA replication, Rep68 and Rep78 regulate the transcription of the AAV p5, p19, and p40 promoters in vivo (BEATON et al.
Virology 181: 738-741 Kotin RM (1994) Prospects for the use of adeno-associated virus as a vector for human gene therapy. Hum Gene Ther 5: 793-801 Kotin RM, Siniscalco M, Samulski RJ, Zhu X, Hunter L, Laughlin CA, McLaughlin S, Muzyczka N, Rocchi M, Berns KI (1990) Site-specific integration by adeno-associated virus. Proc Nat Acad Sci USA 87: 2211-2215 Kotin RM, Menninger JC, Ward DC, Berns KI (1991) Mapping and direct visualization of a regionspecific viral DNA integration site on chrornosome 19q13-qter.
1994). Nested polymerase chain reaction (PCR) amplifications were used to detect AAV-2 DNA in biopsy material from cervix uteri. Larger amounts of AAV-2 DNA were detected using Southern hybridizations of DNA from spontaneous abortion material. In situ hybridization analysis localized the AAV-2 DNA in the villous fraction (trophoblast) of the placenta but not in the embryo or decidua. The presence of AAV-2 DNA in normal uterine tissues suggests that AAV may have established latency at this site.