2004
DOI: 10.1128/jvi.78.20.11263-11271.2004
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Evidence for Gene Expression by Unintegrated Human Immunodeficiency Virus Type 1 DNA Species

Abstract: The integrated form of human immunodeficiency virus type 1 (HIV-1) DNA is classically considered to be the sole template for viral gene expression. However, several studies have suggested that unintegrated viral DNA species could also support transcription. To determine the contribution of the different species of HIV-1 DNA to viral expression, we first monitored intracellular levels of various HIV-1 DNA and RNA species in a single-round infection assay. We observed that, in comparison to the precocity of HIV-… Show more

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Cited by 76 publications
(82 citation statements)
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“…Therefore, reverse transcription and integration are not major contributors to the apparent defectiveness of HIV-1. These results are in general agreement with other published studies using Nef Ϫ HIV-1, HIV-1 vectors, and murine leukemia virus: ϳ10 to 30% of maximum postinfection vDNA become proviruses (3,7,8,11,12,28,43).…”
supporting
confidence: 92%
“…Therefore, reverse transcription and integration are not major contributors to the apparent defectiveness of HIV-1. These results are in general agreement with other published studies using Nef Ϫ HIV-1, HIV-1 vectors, and murine leukemia virus: ϳ10 to 30% of maximum postinfection vDNA become proviruses (3,7,8,11,12,28,43).…”
supporting
confidence: 92%
“…17,18,[22][23][24] Both 1-and 2-LTR episomes can express proteins because the genome or transgene remains intact. 25,26 IN can also auto-integrate within the viral DNA itself, but the disruption of the viral coding region generally renders the resulting products inactive. 17,27 Episomes do not persist in dividing cells because they are not integrated and lack an origin of replication (ORI).…”
Section: Transduction Biologymentioning
confidence: 99%
“…25,61,74,75 In vivo brain injections comparing expression from HIV LV and D64V IDLV found no transgene expression from the IDLV. 74 A later report examined the potential efficacy of HIV D64V IDLV in liver transduction experiments and again no expression was observed.…”
Section: Idlv Biologymentioning
confidence: 99%
“…RNA was prepared from 1 to 2 ϫ 10 6 MCs using the QIAamp RNAeasy Mini Kit protocol (QIAGEN). FL and MS HIV mRNA transcript levels were determined by a modification of a protocol described by Brussels et al 10 Briefly, qRT-PCR was performed in a SYBR Green assay system using a BioRad iCycler programmed for 95°C 3 minutes, (94°C for 1minute, 56°C for 1 minute, 72°C for 1 minutes) ϫ 40 cycles, 4°C for 10 minutes. Primers for FL (sense,: 5Ј-CTGAAGCGCG-CACGGCAA-3Ј, and antisense, 5Ј-GACGCTCTCGCACCCATCTC-3Ј) and for MS sense (GACTCATCAAGTTTCTCTATCAAA) and MS antisense (GTCTCTCAAGCGGTGGT) were used at a final concentration of 200 nM to amplify the products of interest from 400 ng of sample RNA.…”
Section: Analysis Of Hiv ؊ Infection Of Mcs By Real-time Pcrmentioning
confidence: 99%