Term-seq is a service provided by your company that is designed to accurately map 3' ends of bacterial RNA transcripts. This service is particularly useful in bacteria because bacterial mRNAs lack the polyA tail that is present in eukaryotic RNA transcripts, which makes it challenging to map 3' ends using traditional sequencing methods.

Overview of Our Term-Seq

Term-seq works by directly sequencing the exposed RNA 3' ends in bacteria, which allows for the precise mapping of RNA termini in a highly quantitative manner. This provides a genome-wide map of RNA termini, which can be used to better understand the mechanisms of transcription termination in bacteria.

In addition to its utility in mapping RNA termini, Term-seq can also be used to identify novel ribo-regulators, which are RNA molecules that can regulate gene expression. This provides synthetic biologists with additional tools for controlling gene expression in bacteria and can also provide insights into the mechanisms of transcription termination. Overall, Term-seq is a powerful tool for studying gene expression in bacteria and can be used to uncover new regulatory mechanisms and potential therapeutic targets.

Workflow of Our Library Construction

The library construction of Term-seq involves a series of steps that result in the generation of a cDNA library from bacterial RNA transcripts, which is then sequenced using high-throughput sequencing platforms. The resulting sequence reads are processed using bioinformatics tools to accurately map the 3' end termini of RNA transcripts and quantify their expression levels. This provides a highly quantitative and precise map of RNA termini in bacteria, which can be used to study gene expression regulation and transcription termination mechanisms.

Project Workflow

Data analysis

• High-resolution maps of the RNA 3' ends associated with genes
• Research of molecular determinants of RNA 3' ends
• Function of 3' terminal stem-loop structures
• Examine synthetic terminator performance in vivo under a variety of conditions

Deliverable: FastQ, BAM, QC report, Integrative view of the bacterial transcriptome (Multi-layered RNA sequencing data), Folding energy of RNA, Custom bioinformatics analysis.

References:

1. Dar, Daniel, et al. Term-seq reveals abundant ribo-regulation of antibiotics resistance in bacteria. Science, 352.6282 (2016): aad9822.
2. Saliba, Antoine-Emmanuel, et al. New RNA-seq approaches for the study of bacterial pathogens. Current opinion in microbiology, 35 (2017): 78-87.
3. Hudson, Andrew J., et al. Rapid generation of sequence-diverse terminator libraries and their parameterization using quantitative Term-Seq. Synthetic Biology, 4.1 (2019): ysz026.