CRISPR-Cas9 Genome Editing

Nobel prize-winning biochemist Jennifer Doudna
Photo courtesy of Lawrence Berkeley National Laboratory

In October 2020, Jennifer Doudna of UC Berkeley and Lawrence Berkeley National Laboratory and Emmanuelle Charpentier of the Max Planck Unit for the Science of Pathogens in Berlin jointly won the Nobel Prize in Chemistry for the discovery of CRISPR-Cas9, a genome-editing tool that can be programmed to make precise edits to DNA. Doudna’s nascent research on CRISPR RNA strands and the Cas1 protein was funded by DOE’s LDRD program in 2008. Since then, the CRISPR-Cas9 system has been recognized for its vast potential in gene-targeting and gene-editing applications.

Why It Matters

CRISPR-Cas9 genetic engineering technology has profoundly altered genomics research. This genome-editing technology enables scientists to quickly change or remove genes in any organism, including human cells, with unprecedented precision and efficiency. The revolutionary capability and robustness of CRISPR-Cas9 has unlocked sweeping, new possibilities.

Labs worldwide have adapted the course of their research to incorporate this new tool, with huge implications across biology, agriculture, and medicine. In particular, scientists have used CRISPR-Cas enzymes to alter the genetic code of human cells and organs in a way that holds promise to treat genetic disorders, infectious diseases, and cancers. Furthermore, researchers are applying CRISPR-Cas9 to engineer pest- and mold-resistant crops and enhance biofuel production.

Methods

Building upon early findings, Doudna and Charpentier’s research team identified the core mechanisms of the CRISPR-Cas9 system and detailed how it can be programmed to cut DNA at a target sequence. CRISPR, which stands for Clustered Regularly Interspaced Short Palindromic Repeats, is a critical part of the bacterial immune system and handles sequence recognition. Cas9, which stands for CRISPR-assisted, is an RNA-guided enzyme. Cas9 is guided by CRISPR RNA to bind a specific DNA region, and once bound the Cas9 protein subsequently acts like a pair of molecular scissors to accurately cut the DNA strands at the target site. The DNA is cut at a specific location so that it can be edited.

What’s Next

LBNL researchers have moved beyond editing in the cellular genome and developed a technology for making sequence modifications in the mitochondrial genome of eukaryotic cells. These modifications can alter cellular metabolism for fungal, plant, algal and human cells and tissues.

The CRISPR-Cas9 technology is the pioneering basis for many promising medical technologies, including tools to diagnose and treat diseases, and has promising applications for the development of improved crops, biofuels, and bioproducts.

Publications

Martin J, Krzysztof C, Ines F, Michael H, Jennifer D, and Emmanuelle C. 2012. “A Programmable Dual-RNA – Guided DNA Endonuclease in Adaptive Bacterial Immunity.” Science 337(6096): 816-821. DOI: 10.1126/science.1225829.