Enabling Biomanufacturing Through Multiple Microbial Hosts

The Science

DOE’s Joint Genome Institute (JGI) developed a chassis-independent recombinase-assisted genome engineering (CRAGE) tool that allowed inserts from 10 kilo base pairs (Kbp) up to 48 Kbp on nearly 30 gammaproteobacteria species.

The Impact

  • CRAGE saves time as constructs can be inserted in a single step in one day instead of having serial inserts over several days.
  • Novel secondary metabolites not normally produced by the hosts were uncovered. Secondary metabolites are the basis for hundreds of invaluable agricultural, industrial, and medical products.
  • CRAGE can be adapted to other organisms such as fungi and archaea.


A new technology that allows synthetic biologists to domesticate any bacterial host and, in a single step, successfully and efficiently insert constructs.


Wang, G.,  Z. Zhao, J. Ke, Y. Engel, Y.-M. Shi, D. Robinson, K. Bingol, Z. Zhang, B. Bowen, K. Louie, B. Wang, R. Evans, Y.Miyamoto, K. Cheng, S. Kosina, M. De Raad, L. Silva, A. Luhrs, A. Lubbe, D. W. Hoyt, C. Francavilla, H. Otani, S. Deutsch, N. M. Washton, E. M. Rubin, N. J. Mouncey, A. Visel, T. Northen, J.-F. Cheng, H. B. Bode, and Y. Yoshikuni. 2019. “CRAGE Enables Rapid Activation of Biosynthetic Gene Clusters in Undomesticated Bacteria,Nature Microbiology 4, 2498–2510. DOI:10.1038/s41564-019-0573-8.