Using CRISPR Genetic Technology catch cancer in the act

Using CRISPR Genetic Technology catch cancer in the act

Phylogenetic trees, starting with an individual cancer cell. Each color represents a different location in the body. A very colorful tree shows a highly metastatic phenotype, where the descendants of a cell have jumped between different tissues several times. A tree that is predominantly one color represents a less metastatic cell. Credit: Jeffrey Quinn / Whitehead Institute

Using CRISPR technology, researchers track the lineage of individual cancer cells as they proliferate and metastasize in real time.

When cancer is confined to one place in the body, doctors can often treat it with surgery or other therapies. Much of the mortality associated with cancer, however, is due to its tendency to metastasize, sending seeds of itself that can take root throughout the body. The exact moment of metastasis is fleeting, lost in the millions of divisions that take place in a tumor. “These events are generally impossible to monitor in real time,” says Jonathan Weissman, professor of biology at MIT and member of the Whitehead Institute for Biomedical Research.

Now, researchers led by Weissman, who is also a researcher at Howard Hughes Medical Institute, have turned a CRISPR tool into a way to do it. In an article published Jan. 21, 2021 in Science, Weissman’s lab, working with Nir Yosef, a computer scientist at the University of California at Berkeley, and Trever Bivona, a cancer biologist at the University of California at San Francisco, discusses cancer cells like evolutionary biologists could look at species, building a complex and detailed family tree. By examining the branches, they can track the cell’s lineage to find when a single tumor cell has gone rogue, spreading its offspring to the rest of the body.

“With this method, you can ask questions like, ‘How often does this tumor metastasize? Where do metastases come from? Where are they going? “Weissman says. “By being able to follow the history of the tumor in vivo, you reveal differences in tumor biology that were otherwise invisible.”

Scratch paper cells

Scientists have followed cancer cell lines in the past by comparing shared mutations and other variations in their DNA patterns. However, these methods depend to some extent on the existence of enough natural mutations or other markers to accurately show the relationships between cells.

It was there that Weissman and co-first authors Jeffrey Quinn, then a post-doctoral fellow in Weissman’s lab, and Matthew Jones, a graduate student of Weissman’s lab, saw the opportunity to use CRISPR technology – in particular, a method developed by Michelle Chan, member of the Weissman Lab, to track embryo development – to facilitate tracking.

Instead of just hoping that a cancerous line would contain enough lineage-specific markers to follow, the researchers decided to use Chan’s method to add the markers themselves. “Basically, the idea is to design a cell that has a genomic DNA pad, which can then be ‘written’ using CRISPR,” Weissman explains. This “writing” in the genome is done in such a way that it becomes hereditary, which means that the small-offspring of a cell would see the “writing” of its mother cells and its grandparent cells recorded in its. genome.

To create these special “scratchpad” cells, Weissman designed human cancer cells with additional genes: one for the bacterial Cas9 protein – the famous “molecular scissors” used in CRISPR genome editing methods – others for bright proteins for microscopy, and some sequences would serve as targets for CRISPR technology.

They then implanted thousands …

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