Deciphering Disease Progression And Cell Processes With TIGER, In Vivo And Noninvasively
Could prospective patients take a diagnostic ribonucleic acid probiotic designed to test gut health with individual cells? Researchers at the Helmholtz Institute for RNA-Based Infection Research (HIRI) and the Julius Maximilian University of Würzburg (JMU) have developed a new technology called TIGER. This allows deciphering complex processes in individual cells in vivo by recording past RNA transcripts. The results were published in the journal Nature Biotechnology on January 5, 2023.
Bacterial and viral infections can cause acute acute symptoms, but can also have devastating long-term effects, such as cancer. As a result, scientists are looking for new approaches and technologies to better understand the disease process and predict the development of cells and tissues. They used more precise methods to analyze the processes behind individual cells. One of its goals is to detect changes in gene activity, which, in turn, can indicate pathological phenomena.
Ribonucleic acid (RNA) makes an important contribution to this understanding. They can show gene activity because only an active gene makes a copy of the RNA (transcript) in a process called transcription. However, the RNA molecules expressed in the transcript represent only the current state. Relating past cellular events, such as bacterial infection, to current conditions and inferring future outcomes has proven difficult.
"The identity and behavior of a cell depends not only on its current intracellular composition and extracellular environment, but also on its previous state. We were looking for efficient procedures at the cellular level to look at the past and connect it to the present," explains Professor Chase Beisel, Head of RNA Synthetic Biology at the Institute for RNA Infection Research. Helmholtz (HIRI). .) in Würzburg and lead author of the study.
In their publication, the authors present a new technological approach that could significantly improve medical diagnosis in the future. Their method, called TIGER, is a way to detect the presence of specific RNAs in individual living cells.
"Through RNA recording, TIGER links the current cellular state to the previous transcriptional state," said first author Chunlei Jiao. TIGER can measure relative gene expression, detect differences between single nucleotides, record multiple transcripts simultaneously, and read out events in a single cell.
The method has surprising advantages, says Jiao. "Previous research has only been able to estimate the past state of cells, using large amounts of data and computational prediction tools to measure cells asynchronously over time." The scientists involved in this study were able to document the transfer of antibiotic resistance between E. coli cells, as well as the invasion of Salmonella into the host cell.
In the future, TIGER can be used to study the transcriptional history of individual cells in living organisms and correlate it with the current status quo to decipher complex cellular responses in vivo and noninvasively. For example, one could imagine taking TIGER probiotics to further record and analyze the state of the digestive tract, the authors conclude.
TIGER (short for Putative RNA Transcripts of Genetically Encoded Entries) uses reprogrammed tracrRNAs (Rptrs) to record selected cellular transcripts as DNA modifications stored in individual living bacterial cells. Rptr is designed to associate with a recognized transcript and convert it into a control RNA.
The guide RNA then signals the Cas9 base editor to target the introduced target DNA. The main level of changes can be read in order. The technology builds on previous research that led to the development of LEOPARD, an in vitro diagnostic platform.
Additional information: Chunlei Jiao et al., RNA Recording in Single Bacterial Cells Using Reprogrammed tracrRNA, Nature Biotechnology (2023). DOI: 10.1038/s41587-022-01604-8
Acquired . Deciphering disease progression and cellular processes with TIGER, in vivo and non-invasively (Jan 10, 2023). Retrieved January 12, 2023, from https://phys.org/news/2023-01-deciphering-disease-cell-tiger-live. .html:
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