Grant: Discovery Grants Program - Individual

Genome editing, the ability to modify the DNA within cells, is a powerful tool in science, and has made huge strides in recent years. The game-changer was the application of CRISPR-Cas9 to genome editing in 2012, making editing genes easier and more versatile. These contributions were awarded the Nobel Prize in Chemistry to Drs. Jennifer Doudna and Emmanuelle Charpentier in 2022. CRISPR genome editing has been used in many fields like biology, medicine, and agriculture. But the first version had some problems, like unintended changes in the DNA, which all have been related to cuts in both strands of DNA. To make it better, researchers have been working on CRISPR 2.0 tools that can modify DNA but only cut one strand. In the last decade, significant progress has been made with the development of 4 new CRISPR 2.0 tools that do not create double strand breaks. The first two are base editors, which can change a single letter in the genetic code. One type of base editor turns a cytosine (C) to thymine (T), and the other turns an adenine (A) to guanine (G). The third tool is called a prime editor, which can also make precise changes of up to 50 letters in the genetic code. Finally, a system called PASTE now allows for insertion of thousands of letters into the genome. But there's still a problem - how do we check if these edits worked, particularly in living subjects? Right now, we must collect tissues using invasive procedures or sacrifice the subject to collect tissues of interest. We need a way to check editing while the subject is still alive and without having to collect any tissues. My lab is proposing to build imaging tools that let us see edited cells deep inside living subjects. We're using special genes called reporter genes that can be detected outside the body with different imaging tools such as bioluminescence imaging, magnetic resonance imaging and positron emission tomography. Our goal is to create a set of tools to watch genome editing happening inside living things as well as track cells that are edited in a dish in a lab and transferred into the body. We will build and test these tools in cells and then in mice to see if we can really see the edits happening. This research is important because it can help us understand genome editing better. With these tools, we can see what's happening inside an organism as we edit its genes. In simpler terms, we're improving powerful CRISPR-based genome-editing tools, and making it easier to see if the edits work in living things. This can help us better understand and use genome editing in biology, medicine, agriculture, and more.