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Cross-clade inhibition of HIV-1 replication and cytopathology by using RNase P-associated external guide sequences.

Publication ,  Journal Article
Kraus, G; Geffin, R; Spruill, G; Young, AK; Seivright, R; Cardona, D; Burzawa, J; Hnatyszyn, HJ
Published in: Proc Natl Acad Sci U S A
March 19, 2002

RNase P complexes have been proposed as a novel RNA-based gene interference strategy to inhibit gene expression in human malignancies and infectious diseases. This approach is based on the sequence-specific design of an external guide sequence (EGS) RNA molecule that can specifically hybridize to almost any complementary target mRNA and facilitate its cleavage by the RNase P enzyme component. We designed a truncated RNase P-associated EGS molecule to specifically recognize the U5 region of HIV-1 mRNA and mediate cleavage of hybridized mRNA by the RNase P enzyme. Genes encoding for this U5-EGS (560) molecule, as well as a U5 EGS (560D) antisense control, were cloned into retroviral plasmids and transferred into a CD4(+) T cell line. Transfected cells were exposed to increasing concentrations of HIV-1 clinical isolates from clades A, B, C, and F. Heterogeneous cultures of CD4(+) T cells expressing the U5 EGS (560) molecule were observed to maintain CD4 levels, were devoid of cytopathology, and did not produce HIV p24 gag antigen through 30 days after exposure to all HIV-1 clades at a multiplicity of infection of 0.01. Identical cells expressing the U5 EGS (560D) antisense control molecule underwent a loss of CD4 expression, produced elevated levels of HIV-1, and formed large syncytia similar to untreated cells. When the viral inoculum was increased at the time of exposure (multiplicity of infection = 0.05), the inhibitory effect of the U5 EGS (560) molecule was overwhelmed, but viral-mediated cytopathology and particle production were delayed compared with control cell populations. Viral replication and cytopathology associated with infection of multiple HIV-1 clades can be effectively inhibited in CD4(+) cells expressing the RNase P-associated U5 EGS (560) molecule.

Duke Scholars

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

March 19, 2002

Volume

99

Issue

6

Start / End Page

3406 / 3411

Location

United States

Related Subject Headings

  • Virus Replication
  • Transfection
  • Substrate Specificity
  • Ribonuclease P
  • Reverse Transcriptase Polymerase Chain Reaction
  • Retroviridae
  • RNA, Small Untranslated
  • RNA, Catalytic
  • Proviruses
  • Plasmids
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Kraus, G., Geffin, R., Spruill, G., Young, A. K., Seivright, R., Cardona, D., … Hnatyszyn, H. J. (2002). Cross-clade inhibition of HIV-1 replication and cytopathology by using RNase P-associated external guide sequences. Proc Natl Acad Sci U S A, 99(6), 3406–3411. https://doi.org/10.1073/pnas.052651199
Kraus, Gunter, Rebeca Geffin, Gina Spruill, Andrea K. Young, Rachel Seivright, Diana Cardona, Jennifer Burzawa, and H James Hnatyszyn. “Cross-clade inhibition of HIV-1 replication and cytopathology by using RNase P-associated external guide sequences.Proc Natl Acad Sci U S A 99, no. 6 (March 19, 2002): 3406–11. https://doi.org/10.1073/pnas.052651199.
Kraus G, Geffin R, Spruill G, Young AK, Seivright R, Cardona D, et al. Cross-clade inhibition of HIV-1 replication and cytopathology by using RNase P-associated external guide sequences. Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3406–11.
Kraus, Gunter, et al. “Cross-clade inhibition of HIV-1 replication and cytopathology by using RNase P-associated external guide sequences.Proc Natl Acad Sci U S A, vol. 99, no. 6, Mar. 2002, pp. 3406–11. Pubmed, doi:10.1073/pnas.052651199.
Kraus G, Geffin R, Spruill G, Young AK, Seivright R, Cardona D, Burzawa J, Hnatyszyn HJ. Cross-clade inhibition of HIV-1 replication and cytopathology by using RNase P-associated external guide sequences. Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3406–3411.
Journal cover image

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

March 19, 2002

Volume

99

Issue

6

Start / End Page

3406 / 3411

Location

United States

Related Subject Headings

  • Virus Replication
  • Transfection
  • Substrate Specificity
  • Ribonuclease P
  • Reverse Transcriptase Polymerase Chain Reaction
  • Retroviridae
  • RNA, Small Untranslated
  • RNA, Catalytic
  • Proviruses
  • Plasmids