CRISPR tools have been shown to be highly effective at neutralizing RNA viruses in the cytoplasm

Researchers from Helmholtz München and the Technical University of Munich (TUM) have developed a new tool (Cas13d-NCS) that allows CRISPR RNA molecules present inside cells to move into the cytoplasm and effectively neutralize RNA viruses. It is reported that it has been developed. . The research team believes that the technology is necessary because RNA-targeting tools like CRISPR/Cas13 are powerful but inefficient in the cell cytoplasm, where many RNA viruses replicate, and that precision medicine and aggressive virus defense strategies are needed. It says that it will open the door to.

Scientist's researchEngineered nucleocytoplasmic shuttle Cas13d enables highly efficient cytosolic RNA targetingAppears in “ discovery of cells.

“The CRISPR/Cas13 system is a programmable tool for manipulating RNA and has been used in a variety of RNA-targeting applications. Within the Cas13 family, Cas13d is the most active subunit in mammalian cells. type. Recently, Cas13d was utilized as an antiviral drug against various human RNA viruses,” the researchers wrote.

“However, Cas13d has little activity in the cytosol of mammalian cells, and its activity is restricted to the nucleus, limiting its applications as a programmable antiviral drug. Most RNA viruses are found in the cytosol. This suggests that current Cas13d-based antivirals rely on uncontrolled nuclear leakage and are therefore limited in their efficiency.

“Here we show that the nuclear localization of Cas13d crRNA is the underlying cause of Cas13d's nuclear preference. To address this limitation, we synthesized nucleocytoplasmic shuttling Cas13d (Cas13d-NCS). We designed Cas13d-NCS to transport nuclear crRNA to the cytosol, where a protein/crRNA complex binds and degrades the complementary target RNA.

“We screened different designs of shuttle proteins and characterized multiple design parameters for the best-performing systems. demonstrated that it is excellent at degrading derived self-replicating RNA.

“Finally, we utilized Cas13d-NCS to completely block the replication of various SARS-CoV-2 strains. It enables operations such as

Screening and optimization

Through screening and optimization, the researchers developed what they called the “transformation solution, Cas13d-NCS.” This is a system that allows nuclear CRISPR RNA (crRNA) to be imported into the cytosol and direct CRISPR-Cas complexes to specific target sequences for precise modification. In the cytosol, the researchers say, protein/crRNA complexes target complementary RNAs and degrade them with unprecedented precision.

Cas13d-NCS surpasses previous products in degrading mRNA targets and neutralizing self-replicating RNA, including replicating sequences of the Venezuelan equine encephalitis (VEE) RNA virus and some variants of SARS-CoV-2. and maximize the potential of Cas13d as a programmable antiviral drug. Scientists say this result is an important step in fighting the pandemic and strengthening defenses against future outbreaks.

The impact of this research extends beyond traditional antiviral strategies and the CRISPR system, ushering in a new era of precision medicine by enabling strategic manipulation of the subcellular localization of CRISPR-based interventions. said Dr. Wolfgang Wurst, Director of the Institute of Genetics. Helmholtz Munich.

“This breakthrough in antiviral drug development using Cas13d-NCS marks a pivotal moment in the ongoing fight against RNA viruses,” said Coordinator of the study and Director of German Neuroscience says Dr. Wurst, who is also chair of developmental genetics at TUM, a partner in the Center for Degenerative Diseases. DZNE) in Munich. He is a member of the Systems Neurology Cluster (SyNergy). “This achievement demonstrates the power of collaborative innovation and human ingenuity in our quest for a healthier and more resilient world.”