Efficient gene transfer of HIV-1-specific short hairpin RNA into human lymphocytic cells using recombinant adeno-associated virus vectors.
The cellular introduction of short, interfering RNA leads to sequence-specific degradation of homologous mRNA, a process termed RNA interference (RNAi). Here, we report that recombinant adeno-associated virus 2 (rAAV-2) can be used to transfer short hairpin (sh) RNA expression cassettes genetically into human cells. HIV-1 replication was suppressed by >95% in H9 cells and primary human lymphocytes that expressed shRNA targeting the first exon of the viral transactivator protein tat compared to control cells. rAAV-2 integrated stably into the host genome, leading to long-term expression of tat shRNA. Our findings demonstrate the utility of rAAV-2 for the genetic transfer of shRNA expression cassettes into human cells, providing an alternative to using retroviral vectors as RNAi delivery systems.
Boden
Daniel
D
Laboratory of Retrovirology, Division of Infectious Diseases, Department of Medicine, Brown Medical School, Providence, RI 02903, USA.
Pusch
Oliver
O
Lee
Fred
F
Tucker
Lynne
L
Ramratnam
Bharat
B
D015152:Blotting, Northern; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D005091:Exons; D015696:Gene Products, tat / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism* / Q000821:virology*; D008957:Models, Genetic; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012313:RNA / Q000737:chemistry*; D034622:RNA Interference; D012190:Retroviridae / Q000235:genetics; D013997:Time Factors; D015533:Transcriptional Activation; D054322:tat Gene Products, Human Immunodeficiency Virus
D015696:Gene Products, tat; D054322:tat Gene Products, Human Immunodeficiency Virus; D012313:RNA