Reverse transcription of the human immunodeficiency virus type 1 is characterized by the formation of a DNA flap at the center of the viral cDNA in between the central polypurine tract (cPPT) and the central termination sequence (CTS). The importance of the DNA flap for HIV-1 replication has been questioned, whereas its importance for lentiviral vector performance is well accepted. To investigate this controversy, we re-evaluated the importance of the DNA flap for HIV-1 replication. A flap negative HIV-1 virus showed a 10- to 100-fold replication defect in comparison with a WT strain. Further characterization of the DNA flap in the context of lentiviral vectors showed that mutations in the DNA-flap sequence did not affect the transduction efficiency. Finally, introduction of a second cPPT/CTS sequence resulted in the presence of two DNA flaps but no higher transduction efficiency.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.bbrc.2006.08.141 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Flap endonuclease 1 repairs DNA-protein cross-links via ADP-ribosylation-dependent mechanisms.
Sci Adv
January 2025
Developmental Therapeutics Branch, Center for Cancer Research, National Cancer Institute, National Institute of Health, Bethesda, MD 20892, USA.
DNA-protein cross-links (DPCs) are among the most detrimental genomic lesions. They are ubiquitously produced by formaldehyde (FA), and failure to repair FA-induced DPCs blocks chromatin-based processes, leading to neurodegeneration and cancer. The type, structure, and repair of FA-induced DPCs remain largely unknown.
View Article and Find Full Text PDFStructure-switchable branched inhibitors regulate the activity of CRISPR-Cas12a for nucleic acid diagnostics.
Anal Chim Acta
January 2025
Department of Laboratory Medicine, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, People's Republic of China; Wuhan Research Center for Infectious Diseases and Cancer, Chinese Academy of Medical Sciences, Wuhan, People's Republic of China; Hubei Engineering Center for Infectious Disease Prevention, Control and Treatment, Wuhan, People's Republic of China. Electronic address:
Background: In current years, the CRISPR (clustered regularly interspaced short palindromic repeats) based strategies have emerged as the most promising molecular tool in the field of gene editing, intracellular imaging, transcriptional regulation and biosensing. However, the recent CRISPR-based diagnostic technologies still require the incorporation of other amplification strategies (such as polymerase chain reaction) to improve the cis/trans cleavage activity of Cas12a, which complicates the detection workflow and lack of a uniform compatible system to respond to the target in one pot.
Results: To better fully-functioning CRISPR/Cas12a, we reported a novel technique for straightforward nucleic acid detection by incorporating enzyme-responsive steric hindrance-based branched inhibitors with CRISPR/AsCas12a methodology.
DNAzyme assisted single amplification for FEN1 activity detection using a personal glucose meter.
Anal Chim Acta
January 2025
MOE Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, 350116, PR China. Electronic address:
Flap endonuclease 1 (FEN1) plays a vital role in cancer by modulating DNA repair mechanisms, inducing genomic instability, and serving as a promising biomarker for cancer diagnosis and prognosis. In this work, we present the development of a novel DNAzyme signal amplification-directed point-of-care sensing system (Dz-PGM) for the sensitive and specific detection of FEN1. The Dz-PGM system utilizes DNAzyme signal amplification in conjunction with a personal glucose meter (PGM) for reporting, capitalizing on a biochemical cascade initiated by FEN1 recognition.
View Article and Find Full Text PDFApplication and development of biosensing strategies for the analysis of the activity of the tumour-associated enzyme FEN1.
Talanta
January 2025
Department of Transfusion Medicine, West China Hospital of Sichuan University, Sichuan, 610041, PR China. Electronic address:
As a core genetic biomolecule in ecosystems, the metabolic processes of DNA, particularly DNA replication and damage repair, are regulated by Flap endonuclease 1 (FEN1). Abnormal expression and dysfunction of FEN1 may lead to genomic instability, which can induce a variety of chromosome-associated disorders, including tumours. FEN1 has emerged as a prominent tumour marker.
View Article and Find Full Text PDFGold nanocube-enhanced SERS biosensor based on heated electrode coupled with exonuclease III-assisted cycle amplification for sensitive detection of flap endonuclease 1 activity.
Talanta
December 2024
Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian, 350108, China. Electronic address:
The flap endonuclease 1 (FEN1) plays a key role in DNA replication and repair, its aberrant expression is associated with tumor development, so it has been recognized as a promising biomarker for a variety of cancers. Here, a novel "turn on" mode gold nanocube-enhanced surface-enhanced Raman scattering (SERS) biosensor was constructed by combining a heated Au electrode (HAuE), exonuclease III (Exo III)-assisted cycle amplification, and gold nanocube (AuNC)-based SERS enhancement to achieve highly sensitive detection of FEN1 activity. The SERS tag was prepared using the Raman reporter modified on the AuNC surface, and the high electromagnetic field provided by the sharp geometric feature of AuNC greatly enhanced the SERS signal.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!