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Related Concept Videos

Retroviruses02:33

Retroviruses

Retroviruses and retrotransposons both insert copies of their genetic elements into the genome of the host cell. Thus, the viral genes are passed on when the host genome is replicated or translated. A typical retroviral DNA sequence contains 3-4 genes that encode the different proteins required for its structural assembly and function as a molecular parasite. This DNA is transcribed into a single mRNA, which is very similar in structure to conventional mRNAs, i.e., it is capped at the 5’...
Retrovirus Life Cycles01:10

Retrovirus Life Cycles

Retroviruses have a single-stranded RNA genome that undergoes a special form of replication. Once the retrovirus has entered the host cell, an enzyme called reverse transcriptase synthesizes double-stranded DNA from the retroviral RNA genome. This DNA copy of the genome is then integrated into the host’s genome inside the nucleus via an enzyme called integrase. Consequently, the retroviral genome is transcribed into RNA whenever the host’s genome is transcribed, allowing the retrovirus to...
Viruses with RNA Genomes01:29

Viruses with RNA Genomes

RNA viruses are categorized into positive-strand, negative-strand, or double-stranded groups based on their genomic structure and replication mechanisms. This classification dictates how they exploit host cellular machinery for protein synthesis and replication. Some RNA viruses also utilize reverse transcription as part of their life cycle, further diversifying their replication strategies.Positive-Strand RNA VirusesPositive-strand RNA viruses have genomes that function directly as messenger...
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...
DNA Packaging00:58

DNA Packaging

Overview
DNA Packaging00:58

DNA Packaging

Overview

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Related Experiment Video

Updated: May 9, 2026

Packaging HIV- or FIV-based Lentivector Expression Constructs &amp; Transduction of VSV-G Pseudotyped Viral Particles
11:08

Packaging HIV- or FIV-based Lentivector Expression Constructs & Transduction of VSV-G Pseudotyped Viral Particles

Published on: April 8, 2012

Packaging shRNA retroviruses.

Kenneth Chang, Krista Marran, Amy Valentine

    Cold Spring Harbor Protocols
    |August 3, 2013
    PubMed
    Summary
    This summary is machine-generated.

    This study details using viral vectors to deliver short-hairpin RNA (shRNA) for gene silencing in mammalian cells. Optimizing viral titers involves targeting microRNA biogenesis factors like DGCR-8/Pasha.

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    Lentivirus Production

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    Last Updated: May 9, 2026

    Packaging HIV- or FIV-based Lentivector Expression Constructs &amp; Transduction of VSV-G Pseudotyped Viral Particles
    11:08

    Packaging HIV- or FIV-based Lentivector Expression Constructs & Transduction of VSV-G Pseudotyped Viral Particles

    Published on: April 8, 2012

    Designing, Packaging, and Delivery of High Titer CRISPR Retro and Lentiviruses via Stereotaxic Injection
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    Designing, Packaging, and Delivery of High Titer CRISPR Retro and Lentiviruses via Stereotaxic Injection

    Published on: May 23, 2016

    Lentivirus Production
    11:42

    Lentivirus Production

    Published on: October 2, 2009

    Area of Science:

    • Molecular Biology
    • Gene Regulation
    • Virology

    Background:

    • RNA interference (RNAi) is a powerful tool for gene silencing.
    • Viral vectors are commonly used to deliver genetic material into target cells.
    • Short-hairpin RNA (shRNA) is a key component for initiating RNAi.

    Purpose of the Study:

    • To describe the process of using viral vectors for shRNA delivery.
    • To highlight critical considerations for packaging cell line selection and viral host range.
    • To present a method for enhancing viral titers by targeting microRNA biogenesis.

    Main Methods:

    • Integration of shRNA into a stable transgene within the host cell genome.
    • Infection of target cells using viral plasmids containing shRNA.
    • Transfection of plasmids into packaging cell lines for virus production.
    • Selection of appropriate packaging cell systems and envelope proteins (e.g., ecotropic, amphotropic, VSV glycoprotein).
    • Cotransfection with small interfering RNA (siRNA) targeting DGCR-8/Pasha or Drosha to enhance viral titers.

    Main Results:

    • Viral vectors efficiently deliver shRNA for gene silencing.
    • Packaging cell lines and envelope proteins determine viral host range and stability.
    • shRNA cleavage by host RNAi machinery can reduce viral titers.
    • Cotransfection with siRNA targeting DGCR-8/Pasha or Drosha significantly enhances viral titers.

    Conclusions:

    • Viral delivery of shRNA is an effective strategy for mammalian gene silencing.
    • Optimization of packaging and delivery systems is crucial for successful gene silencing experiments.
    • Targeting microRNA biogenesis pathways offers a method to improve the efficiency of viral vector-mediated gene silencing.