Gene silencing of HIV chemokine receptors using ribozymes and single-stranded antisense RNA.
The chemokine receptors CXCR4 and CCR5 are required for HIV-1 to enter cells, and the progression of HIV-1 infection to AIDS involves a switch in the co-receptor usage of the virus from CCR5 to CXCR4. These receptors therefore make attractive candidates for therapeutic intervention, and we have investigated the silencing of their genes by using ribozymes and single-stranded antisense RNAs. In the present study, we demonstrate using ribozymes that a depletion of CXCR4 and CCR5 mRNAs can be achieved simultaneously in human PBMCs (peripheral blood mononuclear cells), cells commonly used by the virus for infection and replication. Ribozyme activity leads to an inhibition of the cell-surface expression of both CCR5 and CXCR4, resulting in a significant inhibition of HIV-1 replication when PBMCs are challenged with the virus. In addition, we show that small single-stranded antisense RNAs can also be used to silence CCR5 and CXCR4 genes when delivered to PBMCs. This silencing is caused by selective degradation of receptor mRNAs.
Qureshi
Amer
A
Randall Division of Cell and Molecular Biophysics, King's College London, University of London, Guy's Campus, London Bridge, London SE1 1UL, UK.
Zheng
Richard
R
Parlett
Terry
T
Shi
Xiaoju
X
Balaraman
Priyadhashini
P
Cheloufi
Sihem
S
Murphy
Brendan
B
Guntermann
Christine
C
Eagles
Peter
P
D002478:Cells, Cultured; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D005434:Flow Cytometry; D015870:Gene Expression; D020868:Gene Silencing; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism / Q000821:virology; D016372:RNA, Antisense / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D016337:RNA, Catalytic / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D019713:Receptors, CCR5 / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication
D016372:RNA, Antisense; D016337:RNA, Catalytic; D012333:RNA, Messenger; D019713:Receptors, CCR5; D019718:Receptors, CXCR4; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases