Tn5 Transposase
Tn5 Transposase - Related Videos
05:20
Hyperactive piggyBac Transposase-mediated Germline Transformation in the Fall Armyworm, Spodoptera frugiperda
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Cited by 8 •
2021
Successful germline transformation in the fall armyworm, Spodoptera frugiperda, was achieved using mRNA of hyperactive piggyBac transposase.
08:15
gDNA Enrichment by a Transposase-based Technology for NGS Analysis of the Whole Sequence of BRCA1, BRCA2, and 9 Genes Involved in DNA Damage Repair
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2014
gDNA enrichment for NGS sequencing is an easy and powerful tool for the study of constitutional mutations. In this article, we present the procedure to analyse simply the complete sequence of 11 genes involved in DNA damage repair.
02:13
Overview of Transposition and Recombination
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2020
Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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.
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...
FAQs
Tn5 transposase recognizes defined sequences at transposon ends and cuts DNA while carrying out the adjoining insertion step. This coupling, known as tagmentation, places adapter sequences directly into the resulting DNA fragments. Because cleavage and adapter addition occur together, the enzyme can convert genomic DNA into material suitable for downstream sequencing-library preparation.
Transposon-end sequences act as recognition features that guide Tn5 transposase to the DNA context it can process. This recognition step connects the enzyme’s cutting activity with adapter insertion, helping generate tagged fragments according to the transposon design. The resulting sequence-marked DNA can then support analysis of genome structure, function, or regulation.
In ATAC-seq, adapter insertion is not interpreted as random coverage across the genome. Tn5 preferentially inserts adapters in regions that are available to regulatory proteins, so the resulting tagged fragments provide a measurement of chromatin accessibility. Researchers can use that pattern to examine regulatory organization and connect accessible genomic regions with gene regulation.
A typical library-preparation workflow processes genomic DNA with engineered Tn5, allowing the enzyme to cleave DNA and insert adapters in the same operation. The tagged fragments then serve as sequencing-library material. This streamlined coupling avoids treating cleavage and adapter addition as separate operations, supporting rapid preparation for genome analysis.
Depending on the assay, Tn5-based methods can support measurements of genome structure and function, chromatin accessibility, mutation patterns, and features related to gene regulation. These outputs come from analyzing where enzyme-generated, adapter-containing fragments occur or from sequencing the prepared material, allowing genome analysis across multiple biological questions.
Tn5-based approaches are valuable because they provide rapid, scalable measurements across genome-analysis questions. In genetics, that efficiency supports studies of accessible chromatin, genome structure and function, mutation patterns, and gene regulation. Their flexibility makes the same enzyme-centered strategy relevant to both sequencing-library preparation and broader analysis of genomic organization.