title: Refinement of lentiviral vector for improved RNA processing and reduced rates of self inactivation repair.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: SA Pathology, North Adelaide, South Australia, Australia. Rachel.Koldej@STJUDE.ORG
authors.forename: Rachel M
authors.initials: RM
authors.lastname: Koldej
authors.forename: Donald S
authors.initials: DS
authors.lastname: Anson
mesh_terms: D002460:Cell Line; D015316:Genetic Therapy; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D026723:Polyadenylation; D022821:RNA Splice Sites; D012367:RNA, Viral / Q000235:genetics
chemical_list: D022821:RNA Splice Sites; D012367:RNA, Viral