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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...
Exon Recombination02:32

Exon Recombination

The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes. 
Exon shuffling follows “splice frame rules.” Each exon has three reading...
Non-LTR Retrotransposons03:18

Non-LTR Retrotransposons

As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
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...

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Reverse Genetics of Orthoflaviviruses: Strategies for Constructing Functional or Infectious cDNA.

Young-Min Lee1

  • 1Department of Animal, Dairy, and Veterinary Sciences, College of Agriculture and Natural Resources, Utah State University, Logan, Utah, USA.

Journal of Medical Virology
|June 10, 2026
PubMed
Summary

Reverse genetics (RG) systems are crucial for RNA virus research. New DNA-launched systems offer alternatives to traditional RNA-launched systems for orthoflaviviruses, overcoming challenges in genetic stability.

Keywords:
flavivirusfunctional cDNAinfectious cDNAorthoflavivirusreverse genetics

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Area of Science:

  • Virology
  • Molecular Biology

Background:

  • Reverse genetics (RG) systems enable precise manipulation of viral genomes from cloned complementary DNA (cDNA).
  • Orthoflaviviruses, including zoonotic arboviruses, pose significant research challenges for RG development.

Purpose of the Study:

  • To summarize and compare RNA- and DNA-launched RG systems for orthoflaviviruses.
  • To highlight technical challenges and provide a framework for selecting appropriate RG approaches.

Main Methods:

  • Review of existing RNA- and DNA-launched reverse genetics systems.
  • Discussion of bacteria-based and bacteria-free cloning strategies.
  • Emphasis on challenges specific to orthoflavivirus genome manipulation.

Main Results:

  • RNA-launched systems are standard, but DNA-launched systems are gaining interest.
  • Instability and toxicity of certain genomic regions hinder full-length clone construction.
  • Bacteria-free methods like Gibson assembly and circular polymerase extension reaction aid clone stability.

Conclusions:

  • Selecting the right RG system is critical for orthoflavivirus research.
  • Overcoming genetic instability is key for successful virus recovery.
  • Advances in cloning strategies facilitate the application of RG to this viral genus.