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Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Homologous Recombination02:31

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
DNA Isolation01:24

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
Size and Structure of Viral Genomes01:26

Size and Structure of Viral Genomes

Viral genomes exhibit remarkable diversity in size, structure, and composition, influencing their replication strategies and interactions with host cells. These genomes consist of either DNA or RNA and may be linear or circular. Additionally, they can be single-stranded or double-stranded, with each configuration affecting how the virus propagates within a host. RNA viruses, for instance, generally have smaller genomes than DNA viruses, a factor that contributes to their high mutation rates and...

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Related Experiment Video

Updated: Jun 27, 2026

Interview: HIV-1 Proviral DNA Excision Using an Evolved Recombinase
10:20

Interview: HIV-1 Proviral DNA Excision Using an Evolved Recombinase

Published on: June 16, 2008

Interview: HIV-1 proviral DNA excision using an evolved recombinase.

Joachim Hauber

    Journal of Visualized Experiments : Jove
    |December 11, 2008
    PubMed
    Summary

    An engineered recombinase, Tre, efficiently removes human immunodeficiency virus type 1 (HIV-1) proviral DNA from infected human cells. This breakthrough offers a potential strategy to target persistent HIV-1 reservoirs, though challenges in delivery and safety remain.

    Area of Science:

    • Molecular Biology
    • Virology
    • Gene Therapy

    Background:

    • Human immunodeficiency virus type 1 (HIV-1) integrates into host cell chromosomes, forming persistent proviruses.
    • Current HIV-1 treatments suppress viral replication but do not eliminate the integrated provirus, risking drug resistance.
    • Targeting integrated HIV-1 is crucial for developing curative therapies.

    Purpose of the Study:

    • To investigate the efficacy of an engineered recombinase, Tre, in excising integrated HIV-1 proviral DNA.
    • To assess the safety and efficiency of Tre recombinase in human cells.

    Main Methods:

    • Production of loxLTR-containing viral pseudotypes for infecting HeLa cells.
    • Transfection of an HIV-1 infected cell line with a plasmid expressing Tre recombinase.

    More Related Videos

    Molecular Evolution of the Tre Recombinase
    12:02

    Molecular Evolution of the Tre Recombinase

    Published on: May 29, 2008

    Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
    09:31

    Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites

    Published on: March 22, 2016

    Related Experiment Videos

    Last Updated: Jun 27, 2026

    Interview: HIV-1 Proviral DNA Excision Using an Evolved Recombinase
    10:20

    Interview: HIV-1 Proviral DNA Excision Using an Evolved Recombinase

    Published on: June 16, 2008

    Molecular Evolution of the Tre Recombinase
    12:02

    Molecular Evolution of the Tre Recombinase

    Published on: May 29, 2008

    Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
    09:31

    Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites

    Published on: March 22, 2016

  • Monitoring of recombinase activity, proviral DNA deletion, and cell viability.

    • Main Results:

    • The engineered Tre recombinase efficiently excised integrated HIV-1 proviral DNA from infected human cells.
    • No significant cytotoxic effects were observed in treated cells.
    • Demonstrated proof of principle for targeting HIV-1 LTR with engineered recombinases.

    Conclusions:

    • Engineered recombinase Tre can effectively excise HIV-1 provirus from host cell genomes.
    • This approach shows promise for targeting latent HIV-1 reservoirs.
    • Further research is needed to address safe and efficient delivery for therapeutic applications.