The first mechanistic details of interference by a CRISPR-Cas system were obtained by molecular analysis of the Escherichia coli system, consisting of a Cascade complex and a Cas3 nuclease [1]. The initial basic insights of the RNA-guided DNA interference mechanism of the type-I CRISPR-Cas system have provided the basis for genome applications, especially for the analogous DNA-targeting systems Cas9 [2] and Cas12 [3]. Apart from the overwhelming diversity of natural CRISPR-associated nucleases [4], also very interesting synthetic variants have been developed during recent years [5]. In this seminar, selected recent developments will be discussed.
References
[1] Stan J.J. Brouns, Matthijs M. Jore, Magnus Lundgren, Edze R. Westra, Rik J.H. Slijkhuis, Ambrosius P.L. Snijders, Mark J. Dickman, Kira S. Makarova, Eugene V. Koonin, John van der Oost, Small CRISPR RNAs guide antiviral defense in prokaryotes. Science 321(5891), 960-964 (2008). https://doi.org/10.1126/science.1159689
[2] Martin Jinek, Krzysztof Chylinski, Ines Fonfara, Michael Hauer, Jennifer A. Doudna, Emmanuelle Charpentier, A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science 337 (6096), .816-821 (2012). https://doi.org/10.1126/science.1225829
[3] Bernd Zetsche, Jonathan S. Gootenberg, Omar O. Abudayyeh, Ian M. Slaymaker, Kira S. Makarova, Patrick Essletzbichler, Sara E. Volz, Julia Joung, John van der Oost, Aviv Regev, Eugene V. Koonin, Feng Zhang, Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell 163(3), 759-771 (2015). http://dx.doi.org/10.1016/j.cell.2015.09.038.
[4] Kira S. Makarova, Yuri I. Wolf, Jaime Iranzo, Sergey A. Shmakov, Omer S. Alkhnbashi, Stan J. J. Brouns, Emmanuelle Charpentier, David Cheng, Daniel H. Haft, Philippe Horvath, Sylvain Moineau, Francisco J. M. Mojica, David Scott, Shiraz A. Shah, Virginijus Siksnys, Michael P. Terns, Česlovas Venclovas, Malcolm F. White, Alexander F. Yakunin, Winston Yan, Feng Zhang, Roger A. Garrett, Rolf Backofen, John van der Oost, Rodolphe Barrangou, Eugene V. Koonin, Evolutionary classification of CRISPR–Cas systems: a burst of class 2 and derived variants. Nature Reviews Microbiology 18(2), 67–83 (2020). https://doi.org/10.1038/s41579-019-0299-x.
[5] Andrew V. Anzalone, Luke W. Koblan, David R. Liu, Genome editing with CRISPR-Cas nucleases, base editors, transposases and prime editors. Nature Biotechnology 38(7), 824-844 (2020). https://doi.org/10.1038/s41587-020-0561-9
John van der Oost is a molecular microbiologist who obtained his PhD at the Free University of Amsterdam (1989). After postdoc positions at Helsinki University and EMBL-Heidelberg he returned to Amsterdam in 1992 with a fellowship of the Royal Netherlands Academy for Arts and Sciences (KNAW). Since 1995 he is group leader of the Bacterial Genetics group in the Laboratory of Microbiology at Wageningen University. Initially the group focused on the discovery of unique metabolic features of Archaea and Bacteria. Apart from biochemical characterization of key enzymes and regulatory proteins, genomics research was initiated of selected hosts and their viral invaders (genomics, transcriptomics and proteomics). When John was appointed Full Professor (2005), a new research line was initiated, addressing microbial host-virus interactions, including CRISPR-Cas and Argonaute (selected grants: NWO-VICI, NWO-TOP (2x), ERC AdG, ERC-PoC). In addition, he is PI of the NWO gravitation project (2017) that aims at Building a Synthetic Cell. He has been coordinator of an EU project and a Marie Curie Research Training Network. John was elected as member of EMBO (2013), the Royal Dutch Academy (KNAW, 2017), the Royal Dutch Society for Science and Humanities (KHMW, 2019), and the Academia Europaea (2019). He is author of >350 peer- reviewed scientific papers, inventor on 33 patent(-application)s, co-founder/advisor of NTrans Technologies (human gene therapy), advisor of SCOPE Biosciences (pathogen diagnostics), and advisor of Hudson River Biotechnology (plant biotech). In 2018 he received the Spinoza prize (NWO), in 2020 the Lennart Philipson Award (EMBL), and in 2022 the Outstanding Achievement Award of the Neth. Soc. Gene & Cell Therapy (NSGCT).
