Patterns of CRISPR/Cas9 activity in plants, animals and microbes.
The CRISPR/Cas9 system and related RNA-guided endonucleases can introduce double-strand breaks (DSBs) at specific sites in the genome, allowing the generation of targeted mutations in one or more genes as well as more complex genomic rearrangements. Modifications of the canonical CRISPR/Cas9 system from Streptococcus pyogenes and the introduction of related systems from other bacteria have increased the diversity of genomic sites that can be targeted, providing greater control over the resolution of DSBs, the targeting efficiency (frequency of on-target mutations), the targeting accuracy (likelihood of off-target mutations) and the type of mutations that are induced. Although much is now known about the principles of CRISPR/Cas9 genome editing, the likelihood of different outcomes is species-dependent and there have been few comparative studies looking at the basis of such diversity. Here we critically analyse the activity of CRISPR/Cas9 and related systems in different plant species and compare the outcomes in animals and microbes to draw broad conclusions about the design principles required for effective genome editing in different organisms. These principles will be important for the commercial development of crops, farm animals, animal disease models and novel microbial strains using CRISPR/Cas9 and other genome-editing tools.
Institute for Molecular Biotechnology, RWTH Aachen University, Aachen, Germany.
Luisa
L
Bortesi
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Changfu
C
Zhu
Institute for Molecular Biotechnology, RWTH Aachen University, Aachen, Germany.
Julia
J
Zischewski
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Lucia
L
Perez
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Ludovic
L
Bassié
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Riad
R
Nadi
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Giobbe
G
Forni
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Sarah Boyd
SB
Lade
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Erika
E
Soto
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Xin
X
Jin
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Vicente
V
Medina
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Gemma
G
Villorbina
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Pilar
P
Muñoz
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Gemma
G
Farré
Institute for Molecular Biotechnology, RWTH Aachen University, Aachen, Germany.|Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.
Rainer
R
Fischer
TRM Ltd, York, UK.
Richard M
RM
Twyman
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.
Teresa
T
Capell
Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain.|ICREA, Catalan Institute for Research and Advanced Studies, Barcelona, Spain.
Paul
P
Christou
Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.
Stefan
S
Schillberg
D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D004720:Endonucleases / Q000235:genetics* / Q000378:metabolism*; D000072669:Gene Editing; D006801:Humans; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D017393:RNA Editing / Q000235:genetics
D004720:Endonucleases
genome editing; mutational signature; off-target mutations; on-target mutations; sgRNA design; site-directed mutagenesis; species-dependent effects