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

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

BMC Biotechnol

SA Pathology, North Adelaide, South Australia, Australia.

Published: October 2009

AI Article Synopsis

  • Lentiviral gene therapy vectors are being enhanced for clinical use by improving their efficiency and safety, focusing on reducing HIV-1 sequence content and minimizing self-repair of the vector.
  • Key modifications include replacing the polyadenylation signal and mutating splice donor sites to limit RNA splicing and vector mobilisation.
  • The study concludes that targeted design changes can improve RNA processing and decrease unwanted vector repair, creating more effective gene therapy tools.

Article 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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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2765960PMC
http://dx.doi.org/10.1186/1472-6750-9-86DOI Listing

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