ViPS Invited Seminar June 2022

Kan Wang, Iowa State University, USA

Date: 15th June 2022

Time: 14:00

Title: Engineering the “Engineer”: CRISPR RNA-guided Integrase system for Agrobacterium genome editing

Location: Remotely via Zoom

Host: Teemu Teeri


Agrobacterium tumefaciens is a soil bacterium that has been employed for plant genetic transformation in the past 40 years. However, our knowledge about this bacterium and our ability to improve the strain is still limited. Here we report the implementation, for the first time, of a CRISPR RNA-guided integrase system for Agrobacterium genome engineering. In this work, we demonstrate that INsertion of Transposable Elements by Guide RNA–Assisted TargEting (INTEGRATE) can efficiently generate DNA insertions to generate targeted gene knockouts. In addition, in conjunction with the Cre-loxP recombination system, we achieved precise deletions of large DNA fragments. This work provides new genetic engineering strategies for Agrobacterium species and their gene functional analyses.

Professor Kan Wang graduated with her BS in Biochemistry from Fudan University in Shanghai, China. She was sponsored first by the Chinese government then by the Rockefeller Foundation to conduct graduate study under the supervision of Drs. Marc Van Montagu (2013 World Food Prize Laureate) and late Jeff Schell in Ghent University, Ghent, Belgium, the first group that discovered and engineered Agrobacterium tumefaciens for plant genetic transformation. Her PhD study and postdoctoral research were focused on the molecular mechanisms of the T-DNA transfer from Agrobacterium tumefaciens to plant cells. She is regarded as one of the pioneers in understanding the mechanism of DNA transfer from Agrobacterium to plants (Wang et al., Cell, 1984; Science 1987), a fundamental knowledge that became the corner stone for building of tools for plant genetic transformation. Dr. Wang spent 7 years in ICI seeds (now Syngenta) as a project leader in genetic transformation of corn and soybean. In 1996, she took a position at Iowa State University and established the first public crop transformation facility, which provides genetic transformation services of corn, soybean, and rice for the research communities. Her current research interests include exploring novel plant genetic transformation and genome editing technologies, investigating CRISPR off-target activities and understanding the functional roles of Agrobacterium non-coding RNAs.

Dr. Wang is a Global Professor of Biotechnology in the Department of Agronomy, and Co-Director of the Crop Bioengineering Center at Iowa State University. Dr. Wang was the winner of 2015 Iowa Women of Innovation Awards for Research Innovation and Leadership for her soybean transformation method. This invention also earned her 2017 Iowa State University Award for Achievement in Intellectual Property. Dr. Wang is a fellow for the Society of In Vitro Biology.

 Read more about Kan's work

Related publications:

Lee, K, Zhang, Y, Kleinstiver, BP, Guo, JA, Aryee, MJ, Miller, J, Malzahn, A, Zarecor, S, Lawrence-Dill, C, Joung, K, Qi, Y, Wang, K.  Activities and specificities of CRISPR-Cas9 and Cas12a nucleases for targeted mutagenesis in maize.  Plant Biotechnology Journal, 17: 362-372 (2019).

Lee, L, Eggenberger, AL, Banakar, R, McCaw, ME, Zhu, H, Main, M, Kang, M, Gelvin, SB, Wang, K.  CRISPR/Cas9-mediated targeted T-DNA integration in rice.  Plant Molecular Biology, 99: 317-328 (2019).

Char, S-N, Neelakandan, AK, Nahampun, H, Frame, B, Main, M, Spalding, MH, Becraft, PW, Meyers, BC, Walbot, V, Wang, K, Yang, B.  An Agrobacterium-delivered CRISPR/Cas9 system for high-frequency targeted mutagenesis in maize.  Plant Biotechnology Journal, 15: 257–268 (2017).