Researchers have found a new way to analyze cell-to-cell communication using nanoscale particles that act as messengers and transporters, thanks to an innovative method utilizing CRISPR gene-editing technology. These particles, called small extracellular vesicles (sEVs), are crucial in disease transmission and have potential for drug delivery. The newly introduced system, referred to as CIBER, allows for the examination of thousands of genes simultaneously by marking sEVs with a unique RNA “barcode.” This breakthrough aims to uncover the factors that influence the release of sEVs from host cells, thereby enhancing our understanding of sEV biology and contributing to the development of novel treatments for various illnesses, including cancer.
Your body communicates in multiple ways. Cells interact with one another, ensuring that your body parts work together harmoniously. Yet, many aspects of this communication remain unclear. Extracellular vesicles (EVs), tiny particles emitted by cells, were once considered mere waste. However, over recent decades, they have been recognized as essential components due to their links with several diseases, such as cancer, neurodegenerative disorders, and age-related conditions.
Small EVs are vital for communication between cells. Depending on the “cargo” they carry from their parent cells—including RNA, proteins, and lipids—sEVs can support normal tissue function or contribute to disease progression. Due to this complexity, researchers are eager to understand the formation and release of sEVs. Unfortunately, traditional methods for isolating sEVs from other substances and pinpointing the elements that trigger their release are often challenging and labor-intensive. To address this issue, a team in Japan has pioneered a new approach.
“We have created an advanced high-throughput screening tool called CIBER (CRISPR-assisted individually barcoded sEV-based release regulator). This system enables a researcher to conduct a genome-wide screening for sEV release regulators within a matter of weeks to a couple of months, making it significantly more efficient than previous methods,” stated Associate Professor Ryosuke Kojima from the Graduate School of Medicine at the University of Tokyo. “CIBER is expected to be an invaluable resource for in-depth investigation into the generation, release, and variety of sEVs.”
The CIBER method employs CRISPR-guide RNA (gRNA) to disable a particular gene in each cell, which then gets barcoded into the sEVs produced by the cell. This process allows researchers to monitor and evaluate the volume of sEVs released by the host cell. Traditional techniques require separating cells into individual wells, altering the expression and activity of a gene in each well, and subsequently measuring the resulting sEV output. In contrast, the CIBER system permits the simultaneous study of thousands of cells with different genes knocked out, all analyzed in a single pool. This allows scientists to investigate the various intricate factors that influence sEV release and to estimate the quantity and diversity of sEVs generated by each cell.
“Looking ahead, CIBER screening could be utilized to identify therapeutic targets related to sEV release or to boost the production of sEVs for medicinal applications, including cancer therapy,” noted Kojima. “Barcoded sEVs may also serve to assess cell population dynamics non-invasively, and tracking the paths of barcoded sEVs could deepen our understanding of sEV biology. We believe CIBER screening holds significant promise.”
