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Tn5 Transposase, an enzyme from Escherichia coli's transposon Tn5, aids DNA segment relocation within genomes. It recognizes and cleaves transposon ends, facilitating insertion into new genomic locations (transposition). This enzyme is pivotal in genome engineering and genetics, enabling site-specific gene insertion and mutagenesis. Creative Biogene offers robust Tn5 transposase for accelerated research, especially in tagmentation-based sequencing library construction.
Key Features of Our Tn5 Transposase
Tn5 Transposase, a member of the RNase superfamily, is utilized in ATAC-seq to analyze "open" DNA chromatin regions. This highly active mutant inserts sequencing adapters into open chromatin regions, enhancing efficiency compared to natural enzyme levels. During "tagmentation." Tn5 cuts double-stranded DNA and tags fragments with sequencing adapters. Purified and PCR-amplified DNA fragments undergo high-throughput sequencing. Sequencing reads infer chromatin accessibility, transcription factor binding sites, and nucleosome positioning. The number of reads obtained correlates with chromatin openness at single-nucleotide resolution. In addition to its role in ATAC-seq, Tn5 facilitates DNA transposition, enabling antibiotic resistance and serving as a model for protein-DNA interactions in genetic experimentation and genome sequencing.
Case Study 1
This study addresses challenges in identifying open chromatin regions and transcription factor binding sites (TFBs) in filamentous fungi by developing an ATAC-seq platform tailored to these organisms, thereby facilitating the generation of comprehensive ATAC-seq libraries for Aspergillus niger and Aspergillus oryzae under diverse growth conditions. Researchers utilized the Tn5 Transposase in ATAC-seq to overcome challenges in identifying open chromatin regions and transcription factor binding sites (TFBs) in filamentous fungi. This technique allowed high-resolution measurements of chromatin accessibility in Aspergillus niger and Aspergillus oryzae under various conditions. By delineating syntenic orthologues and differentially changed chromatin accessibility regions, researchers revealed conservative functions across species and differential gene expression programs. Importantly, they proposed criteria to determine TFBs, construct data libraries, and identify novel TFBs through in vivo genomic footprint analysis. This comprehensive TFBs map enhances understanding of transcriptional regulation in filamentous fungi.
Figure 1. Researchers utilized Tn5 Transposase to develop an ATAC-seq protocol for Aspergillus species, aiming to identify chromatin accessibility and regulatory elements. (Huang L, et al., 2021)
Case Study 2
Researchers utilized the Tn5 Transposase in the Assay for Transposase Accessible Chromatin (ATAC-seq) to annotate and characterize regulatory elements in pigs and cattle. This method aimed to address the dearth of functional annotation in noncoding regions of livestock genomes, hindering agricultural research progress. The study identified over 300,000 active regulatory elements in pigs and cattle, with a significant proportion being tissue-specific. The predominant motif observed across tissues was the insulator CTCF, indicating its role in chromatin organization. Comparison with mouse data revealed the evolutionary conservation of regulatory elements, highlighting the necessity of species-specific annotation. This comprehensive atlas of chromatin accessibility serves as a crucial resource for elucidating the regulatory landscape in livestock genomes.
Figure 2. Researchers profiled genome-wide chromatin accessibility in eight tissues from adult male Hereford cattle and Yorkshire pigs using modified ATAC-seq, emphasizing Tn5 Transposase's utility. (Halstead MM, et al., 2020)
Case Study 3
Numerous chromatin characteristics are pivotal in governing gene expression. Researchers employ the Tn5 Transposase within the Cleavage Under Targets and Tagmentation (CUT&Tag) method to efficiently generate high-resolution sequencing libraries for profiling diverse chromatin components. In this technique, specific chromatin proteins are bound in situ by antibodies, which then tether a protein A-Tn5 transposase fusion protein. Activation of the transposase facilitates the generation of fragment libraries with high resolution and minimal background noise. This streamlined process, conducted in a single tube or microwell, enables rapid sample preparation from live cells to sequencing-ready libraries within a single day. CUT&Tag proves invaluable for profiling histone modifications, RNA Polymerase II, and transcription factors even with limited cell numbers or single cells.
Figure 3. Tn5 Transposase is employed to augment the tethering of adapters at chromatin protein binding sites in CUT&Tag chromatin profiling. This enables precise genomic fragment enrichment via in situ tethering. (Kaya-Okur HS, et al., 2019)
Q: What is the specificity of the Tn5 Transposase?
A: Tn5 Transposase exhibits high specificity for recognizing and cleaving specific sequences at the ends of the transposon, facilitating precise insertion into the target site within the host genome. This specificity ensures accurate transposition events and minimizes off-target effects.
Q: How efficient is the Tn5 Transposase in fragmenting DNA for library construction?
A: The Tn5 Transposase offered by Creative Biogene is a hyperactive variant, ensuring high efficiency in fragmenting double-stranded DNA during library preparation. This hyperactivity enables rapid and efficient tagmentation, resulting in the generation of sequencing libraries in a short time frame.
Q: Can the Tn5 Transposase be used in various sequencing applications?
A: Yes, the Tn5 Transposase provided by Creative Biogene is versatile and can be employed in various sequencing applications, including ATAC-seq and other tagmentation-based sequencing methods. Its compatibility with different experimental protocols makes it suitable for a wide range of genomics research studies.
Q: What quality control measures are in place to ensure the reliability of the Tn5 Transposase?
A: Our Tn5 Transposase undergoes rigorous quality control measures to ensure its reliability and performance consistency. These measures include sequence verification, purity assessment, and functional validation to guarantee the highest quality product for our customers.
Q: Is the Tn5 Transposase compatible with different DNA samples and concentrations?
A: Yes, the Tn5 Transposase is compatible with various types of DNA samples and concentrations commonly encountered in genomics research. Whether working with genomic DNA, plasmids, or other DNA fragments, our Tn5 Transposase offers flexibility and robust performance across different sample types and concentrations.
| Cat.No. | Product Name | Price |
|---|---|---|
| EMQZ1422 | Robust Tn5 Transposase | Inquiry |
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