Sourcing some ingredients closer to home can be a good practice not only in the produce section but also in synthetic biology labs.
To minimize the use of components derived from viral sources, Rice University bioengineers use human-derived proteins known as mechanosensitive transcription factors to activate silent or poorly expressed genes. We have developed a tool to do this. This allows our cells to naturally turn on certain genes in response to mechanical cues. .
The tool, called CRISPR-DREAM (CRISPR-dCas9 Recruited Enhancement Activation Module, or DREAM), is smaller, more effective, and more medically effective than other cutting-edge technologies used to control gene expression. Low toxicity to useful cell types. According to a study published in Nature Methods.
DREAM tools provide better, safer gene and cell therapies and more precise treatments to address haploinsufficiency disorders, which cause many difficult-to-treat conditions, including epilepsy, some cancers, immune deficiencies, and Alzheimer's disease. It has the potential to enable disease models.
“Many human diseases are caused by the problem of producing too few genes,” said Isaac Hilton, assistant professor of bioengineering and biological sciences at Rice University. “Unfortunately, when we encounter health problems where we don't produce enough of a particular protein or gene product, there are often very few treatment options.”
To address this issue, researchers have recently developed cutting-edge synthetic transcription factors using CRISPR-based targeting systems. Most of these tools are constructed using materials of non-human origin and may come with unwanted side effects.
“One of the challenges with some of the current technologies is that they are constructed using viral proteins that have evolved to reprogram the way our cells work, and the methods are not necessarily beneficial. It's not limited to that,” Hilton said. “And while viral-derived elements can be manipulated to favor host cells, we and other researchers have found that they can still cause some toxicity in human cells. I am observing.”
Rather than relying on proteins from viruses, Varun Mahata, a postdoctoral fellow in Hilton's lab, wanted to use transcription factors that human cells already produce and use. Mahata fused specific parts of these proteins involved in gene activation to a CRISPR-based programmable delivery platform using a method aimed at increasing their transcriptional capacity.
“We took advantage of the natural ability of human-derived transcription factors, proteins responsible for gene synthesis within the cells of our bodies,” said Mahata, lead author of the study. “The transcription activation units we have constructed work with great precision. They are able to induce the gene activity we target and activate transcription very quickly and robustly. ”
They also help address another shortcoming of current synthetic gene activation platforms, which are often too large to be efficiently delivered to human cells.
“What we did was look for small segments of human proteins that could be exploited to apply these techniques more effectively to human cells,” said the study's corresponding author, Texas said Hilton, an academic at the Cancer Prevention Research Institute (CPRIT). “When we started this project to construct synthetic transcription factors using human-derived proteins, we wanted to identify the ideal source material, with compactness as one of the key factors.”
Human mechanosensitive transcription factors, proteins used by our cells and organs to respond to mechanical forces, met the researchers' criteria. In addition to being relatively small, it is fast-acting and widely used in almost all types of human cells.
Additionally, the team was able to simultaneously activate up to 16 different locations on the genome. This is a record number of synthetic transcription factors.
“The reason that ability is especially important is because when our cells perform a function, they don't just turn on a single gene,” Hilton said. “Instead, they typically turn on entire constellations or networks of genes in unison. And using these synthetic transcription factors, they can mimic and manipulate what cells do naturally. became.”
As a proof-of-concept exercise, researchers showed that they can use the DREAM tool to convert skin cells into induced pluripotent stem cells (iPSCs) in a petri dish. Hilton says this feat will have “tremendous biomedical utility” in the long term.
This research was supported by the Cancer Prevention Institute of Texas (RR170030), the National Institutes of Health (R35GM143532, R21EB030772, R56HG012206), the American Heart Association (917025), the Fulbright Program, and the National Council on Science and Technology. Mexico and the National Science Foundation.
- Peer-reviewed research:
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“A compactly designed human mechanosensitive transcriptional activation module enables powerful and versatile synthetic transcriptional control.” | Nature Methods | DOI: 10.1038/s41592-023-02036-1
Authors: Barun Mahata, Alan Cabrera, Daniel Brenner, Rosa Selenia Guerra-Resendez, Jing Li, Jacob Goell, Kaiyuan Wang, Yannie Guo, Mario Escobar, Abinand Krishna Parthasarathy, Hailey Szadowski, Guy Bedford, Daniel Reed, Sunghwan Kim, Isaac B .Hilton
https://www.nature.com/articles/s41592-023-02036-1
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https://news-network.rice.edu/news/files/2023/10/BM_IBH_6a.jpg
Photo Caption: Isaac Hilton (right) is an assistant professor of bioengineering and biological sciences and a CPRIT scholar. Barun Mahata (left) is a postdoctoral fellow in the Hilton lab at Rice University. (Photo credit: Hilton Lab/Amaury F. Bitar Yepes)https://news-network.rice.edu/news/files/2023/10/Figure-for-Nat-Methods-Press-Release_10.4.23.jpg
Photo Caption: The CRISPR-DREAM platform enables new opportunities in synthetic biology, gene and cell therapy. 1) Reprogramming specialized cells (such as skin cells) into induced pluripotent stem cells. 2) Streamline gene therapy vectors for activating genes in clinical settings. 3) Activate poorly expressed genes to help treat various diseases. 4) Restore the dysfunctional immune protective function of her T cells. (Image provided by Hilton Institute/Rice University)
- Related article:
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Synthesis tools send messages from station to station within the DNA.
https://news.rice.edu/news/2022/synthetic-tools-conduct-messages-station-station-dnaNew CRISPR technology targets the complex code of the human genome.
https://news.rice.edu/news/2021/new-crispr-tech-targets-human-genomes-complex-code - Link:
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Hilton Lab: https://hiltonlab.rice.edu
Cancer Prevention Institute of Texas: https://www.cprit.state.tx.us/
Department of Bioengineering: https://bioengineering.rice.edu
George R. Brown School of Engineering: https://engineering.rice.edu/
- About rice:
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Located on a 300-acre forested campus in Houston, Rice University is consistently ranked among the nation's top 20 universities by U.S. News & World Report. Rice University is highly regarded for its schools of architecture, business, continuing studies, engineering, humanities, music, natural sciences, and social sciences, and is also home to the Baker Institute for Public Policy. Rice University has 4,552 undergraduate students and 3,998 graduate students, with an undergraduate to faculty ratio of just under 6:1. Its boarding college system creates close-knit communities and lifelong friendships. This is one reason Rice University is ranked #1 for racial and class interaction and #4 for quality of life by the Princeton Review. Rice University is also rated the best value among private universities by Kiplinger Personal Finance.
