Sylvester researchers understanding Cancer Center, which is part of the University of Miami Miller School of Medicine, have documented their use of new RNA sequencing technology to discover molecular cellular differentiation engines that could lead to better regenerative therapies.
In addition to being used in the laboratory, technique, rapid precision precision sequencing (RPRO-SEQ), has the potential to help doctors understand the pathological states of patients and the response to treatment in real time.
The results appear in two articles published in Molecular cell. The first article was published on July 24, 2025 and the second on August 7, 2025.
We have seen a major bottleneck in the field of emerging RNA profiling. “”
Pradeep Kumar Reddy Cingaram, PHD, first author of the study and deputy scientist, Sylvester understanding Cancer Center, Miller School of Medicine, University of Miami
“The existing methods, although powerful, are simply too slow and require large amounts of biological material. Imagine needing tens of millions of cells and several days just to start, which immediately excluded crucial research on rare cell types or precious biopsies of patients, “he said.
Tester RPRO-SEQ
In the first study, the team used RPRO-SEQ to study the role of a protein complex called integrator in the regulation of gene expression, which was previously not found with the nascent RNA sequencing.
“Ints11, the catalytic subunit of the integrator complex, was a convincing choice for us because we already knew that it was a key player in gene regulation,” said Cingaram.
By using cell reprogramming models to induce neuronal differentiation, they found that when INTS11 was removed from neural cells, the activity of the brain development had changed considerably. The genes that were to be active to prevent neurodevelopmental and psychiatric disorders were disabled when scientists have removed int11.
“The RPRO-SEQ has enabled us to locate an essential role for the INTS11 protein as a regulator of the genes involved in neurodevelopmental disorders in neural cells,” said Ramin Shiekhattar, Ph.D., main author of the study division, the co-leaders of cancer of the cancer program of cancer and Eugenia J.
The technique only required 12 hours and 5,000 cells. Existing technologies need several days and millions of cells. In addition, scientists emphasize that RPRO-SEQ allowed them to understand not only when the genes were on and disabled, but how. “In other words, RPRO-SEQ allows a mechanistic understanding of gene expression changes,” said Shiekhattar.
“The standard RNA sequencing examines RNA” in the equilibrium state ” – the accumulation of what has been done. It’s like seeing how many cars are on the road. But RPro-Seq reveals the arnan `nasing ”-what is done right away. It’s like watching cars leave the factory. This gives us crucial and real -time information on the active transcription of the genes, “said Cingaram.
INTS11 in regenerative medicine
Then, the team used RPRO-SEQ to study the role that INTS11 plays in the capacity of stem cells to differentiate itself in any type of cell in the body, a characteristic called pluripotency. They found that the protein began to play an essential role in the regulation of genes from the second day of embryonic development.
“This considerably widens our understanding of the way in which multiipotence and differentiation are harmonized at the molecular level,” said Shiekhattar.
Using a laboratory model, they found that the integrator complex binds and regulates critical genes for the identity of essential stem cells to maintain pluripotence. In addition, “work is the paradigm shift because it places the integrator complex at the first stages of the transcriptional cycle, called” initiation “, forcing a revision of current theories for transcriptional initiation,” said Shiekhattar.
Although the two results concerning the role of INTS11 in early development are major stages in understanding cell differentiation, scientists emphasize that RPRO-SEQ could be used for many other research and clinical ends, such as sampling of a tumor to see how it reacts to therapy.
“We are eager to apply RPRO-SEQ to a wider range of human clinical samples.
“Overall, RPRO-SEQ could emerge as a precious tool for research and clinical environments, expanding the scope of transcriptomic analyzes and allowing more precise individual health care,” added Shiekhattar.