Transposon-based Transgenesis
Transposon-based Transgenesis - Related Videos
05:40
Tol2 Transposon-Mediated Zebrafish Transgenesis: A Procedure to Generate Transgenic Zebrafish Following the Coinjection of Tol2 System Into Fertilized Zebrafish Embryos
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2025
In this video, we demonstrate a procedure to generate a stable zebrafish transgenic line constitutively expressing a fluorescent voltage indicator protein following the co-injection of the Tol2 transposon system comprising the transposase mRNA and Tol2 transposon construct encoding the fluorescent voltage indicator.
02:57
DNA-only Transposons
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2020
DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure. The donor site from where the transposon is excised is either degraded or...
03:29
PiggyBac Transposon-Mediated Gene Editing in Human iPSCs: A Procedure to Integrate Gene of Interest in Human iPSCs Using PiggyBac Transposon System
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2025
This video demonstrates the procedure to generate motor neurons by transfection of human iPSCs with PiggyBac transposon system by the ectopic expression of lineage-specific transcription factors.
07:23
Mosaic Zebrafish Transgenesis for Evaluating Enhancer Sequences
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Cited by 6 •
2010
We demonstrate our approach to finding potential enhancer elements from developmentally regulated genes and evaluating their function through mosaic zebrafish transgenesis.
08:51
Microinjection for Transgenesis and Genome Editing in Threespine Sticklebacks
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Cited by 30 •
2016
Transgenic manipulations and genome editing are critical for functionally testing the roles of genes and cis-regulatory elements. Here a detailed microinjection protocol for the generation of genomic modifications (including Tol2-mediated fluorescent reporter transgene constructs, TALENs, and CRISPRs) is presented for the emergent model fish, the threespine stickleback.
FAQs
The transposase recognizes terminal DNA sequences that flank the foreign gene in a donor construct. It excises the transposon, including its cargo, and inserts that unit into genomic DNA. This integration allows the introduced sequence to remain associated with the genome, supporting stable expression and inheritance in the engineered cell or organism.
The genomic location of an inserted transposon can influence how the cargo behaves and which genes are affected. An insertion may alter nearby genetic activity or produce different expression outcomes depending on its position. Consequently, researchers must consider insertion site when linking an observed phenotype to the introduced gene or to an insertional mutation.
Engineered transposons can expose cancer-driving genes by altering genomic function at their integration sites. Insertional mutagenesis screens may activate oncogenes, which promote tumor development, or disrupt tumor-suppressor genes, which normally restrain it. The resulting genetic changes help reveal pathways that contribute to cancer initiation and progression.
The number of integrated transgene copies can influence the resulting expression pattern and biological effect. Different copy numbers may therefore produce different phenotypes even when the same cargo is introduced. In cancer studies, copy number should be considered alongside insertion site so researchers do not attribute variation solely to the identity of the candidate gene.
A typical design places the gene of interest between terminal sequences recognized by the transposase in a donor construct. The transposase is then used to excise and integrate that transposon into genomic DNA. Researchers can subsequently assess stable expression and examine how the introduced cargo or resulting genomic changes affect the studied cell or organism.
The approach is useful when researchers need reproducible cancer models carrying defined engineered genetic changes. Integrated transposons can support stable expression of selected genes and can also generate mutations that activate oncogenes or disrupt tumor-suppressor genes. These models help connect specific genetic alterations with tumor-related biology and provide systems for studying cancer development.
Researchers can introduce a candidate gene through an engineered transposon and then examine the biological consequences of its stable expression in a cancer-related model. Comparing the resulting behavior with appropriate engineered controls can indicate whether the gene contributes to relevant cancer phenotypes. Interpretation should account for integration site and copy-number effects.