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    Refinement of lentiviral vector for improved RNA processing and reduced rates of self inactivation repair.

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    Author
    Koldej, RM; Anson, DS
    Date
    2009-10-07
    Source Title
    BMC Biotechnology
    Publisher
    Springer Science and Business Media LLC
    University of Melbourne Author/s
    Koldej, Rachel
    Affiliation
    Medicine and Radiology
    Metadata
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    Document Type
    Journal Article
    Citations
    Koldej, R. M. & Anson, D. S. (2009). Refinement of lentiviral vector for improved RNA processing and reduced rates of self inactivation repair.. BMC Biotechnol, 9 (1), pp.86-. https://doi.org/10.1186/1472-6750-9-86.
    Access Status
    Open Access
    URI
    http://hdl.handle.net/11343/257790
    DOI
    10.1186/1472-6750-9-86
    Open Access at PMC
    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2765960
    Abstract
    BACKGROUND: Lentiviral gene therapy vectors are now finding clinical application. In order to fully exploit their potential it is important that vectors are made as efficient and as safe as possible. Accordingly, we have modified a previously reported vector to improve RNA processing, minimise Human Immunodeficiency Virus Type-1 (HIV-1) sequence content and reduce repair of the self inactivating (SIN) deletion. RESULTS: HIV-1 sequence in the vector was reduced by substituting the polyadenylation signal with a heterologous signal. Mutation of splice donor sites was undertaken to prevent the majority of splicing within the vector genomic RNA. In addition, a number of other sequences within the vector were deleted. The combination of these modifications was able to significantly reduce the rates of both vector mobilisation and repair of the self inactivating deletion after two rounds of marker rescue. CONCLUSION: RNA processing can be improved by mutation of the major and minor HIV-1 splice donor sites in the vector. In addition the rate of vector mobilisation and repair of SIN vectors can be successfully reduced by careful vector design that reduces homology between the 5' and 3' long terminal repeats (LTRs) to a minimum.

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