TY - JOUR
T1 - Activity and specificity of TRV-mediated gene editing in plants
AU - Ali, Zahir
AU - Abulfaraj, Aala A.
AU - Piatek, Marek J.
AU - Mahfouz, Magdy M.
N1 - KAUST Repository Item: Exported on 2020-10-01
Acknowledgements: We would like to thank members of the laboratory for genome engineering at KAUST for helpful discussions and comments. This study is supported by King Abdullah University of Science and Technology (KAUST).
This publication acknowledges KAUST support, but has no KAUST affiliated authors.
PY - 2015/6/3
Y1 - 2015/6/3
N2 - © 2015 Taylor and Francis Group, LLC. Plant trait engineering requires efficient targeted genome-editing technologies. Clustered regularly interspaced palindromic repeats (CRISPRs)/ CRISPR associated (Cas) type II system is used for targeted genome-editing applications across eukaryotic species including plants. Delivery of genome engineering reagents and recovery of mutants remain challenging tasks for in planta applications. Recently, we reported the development of Tobacco rattle virus (TRV)-mediated genome editing in Nicotiana benthamiana. TRV infects the growing points and possesses small genome size; which facilitate cloning, multiplexing, and agroinfections. Here, we report on the persistent activity and specificity of the TRV-mediated CRISPR/Cas9 system for targeted modification of the Nicotiana benthamiana genome. Our data reveal the persistence of the TRVmediated Cas9 activity for up to 30 d post-agroinefection. Further, our data indicate that TRV-mediated genome editing exhibited no off-target activities at potential off-targets indicating the precision of the system for plant genome engineering. Taken together, our data establish the feasibility and exciting possibilities of using virus-mediated CRISPR/Cas9 for targeted engineering of plant genomes.
AB - © 2015 Taylor and Francis Group, LLC. Plant trait engineering requires efficient targeted genome-editing technologies. Clustered regularly interspaced palindromic repeats (CRISPRs)/ CRISPR associated (Cas) type II system is used for targeted genome-editing applications across eukaryotic species including plants. Delivery of genome engineering reagents and recovery of mutants remain challenging tasks for in planta applications. Recently, we reported the development of Tobacco rattle virus (TRV)-mediated genome editing in Nicotiana benthamiana. TRV infects the growing points and possesses small genome size; which facilitate cloning, multiplexing, and agroinfections. Here, we report on the persistent activity and specificity of the TRV-mediated CRISPR/Cas9 system for targeted modification of the Nicotiana benthamiana genome. Our data reveal the persistence of the TRVmediated Cas9 activity for up to 30 d post-agroinefection. Further, our data indicate that TRV-mediated genome editing exhibited no off-target activities at potential off-targets indicating the precision of the system for plant genome engineering. Taken together, our data establish the feasibility and exciting possibilities of using virus-mediated CRISPR/Cas9 for targeted engineering of plant genomes.
UR - http://hdl.handle.net/10754/597449
UR - https://www.tandfonline.com/doi/full/10.1080/15592324.2015.1044191
UR - http://www.scopus.com/inward/record.url?scp=84949935802&partnerID=8YFLogxK
U2 - 10.1080/15592324.2015.1044191
DO - 10.1080/15592324.2015.1044191
M3 - Article
C2 - 26039254
AN - SCOPUS:84949935802
VL - 10
SP - e1044191
JO - Plant Signaling & Behavior
JF - Plant Signaling & Behavior
SN - 1559-2324
IS - 10
ER -