In planta gene targeting can be enhanced by the use of CRISPR/Cas12a

The Plant Journal : for Cell and Molecular Biology
Felix Wolter, Holger Puchta

Abstract

The controlled change of plant genomes by homologous recombination (HR) is still difficult to achieve. We previously developed the in planta gene targeting (ipGT) technology which depends on the simultaneous activation of the target locus by a double-strand break and the excision of the target vector. Whereas the use of SpCas9 resulted in low ipGT frequencies in Arabidopsis, we were recently able to improve the efficiency by using egg cell-specific expression of the potent but less broadly applicable SaCas9 nuclease. In this study, we now tested whether we could improve ipGT further, by either performing it in cells with enhanced intrachromosomal HR efficiencies or by the use of Cas12a, a different kind of CRISPR/Cas nuclease with an alternative cutting mechanism. We could show before that plants possess three kinds of DNA ATPase complexes, which all lead to instabilities of homologous genomic repeats if lost by mutation. As these proteins act in independent pathways, we tested ipGT in double mutants in which intrachromosomal HR is enhanced 20-80-fold. However, we were not able to obtain higher ipGT frequencies, indicating that mechanisms for gene targeting (GT) and chromosomal repeat-induced HR differ. However, using LbCas12a,...Continue Reading

References

May 14, 1996·Proceedings of the National Academy of Sciences of the United States of America·H PuchtaB Hohn
Mar 9, 1999·The Plant Journal : for Cell and Molecular Biology·S J Clough, A F Bent
Feb 24, 2001·Cellular and Molecular Life Sciences : CMLS·C Gutierrez
Jan 27, 2004·Veterinary Microbiology·Crisanto GutierrezRiccardo Missich
Nov 30, 2005·The New Phytologist·Clifford M Bray, Christopher E West
Nov 15, 2007·Proceedings of the National Academy of Sciences of the United States of America·Frank HartungHolger Puchta
Oct 30, 2008·Cell·Louise J BarberSimon J Boulton
Mar 6, 2010·Science·Jillian L YoudsSimon J Boulton
May 29, 2010·Proceedings of the National Academy of Sciences of the United States of America·Feng ZhangDaniel F Voytas
Nov 18, 2010·Journal of Experimental Botany·Alexander Knoll, Holger Puchta
Apr 25, 2012·Proceedings of the National Academy of Sciences of the United States of America·Friedrich FauserHolger Puchta
Jun 23, 2012·Science·Wayne CrismaniRaphaël Mercier
Jun 30, 2012·Science·Martin JinekEmmanuelle Charpentier
Jan 5, 2013·Science·Le CongFeng Zhang
Oct 30, 2013·The International Journal of Developmental Biology·Holger Puchta, Friedrich Fauser
Dec 18, 2013·Pest Management Science·Qin Yu, Stephen B Powles
Jan 21, 2014·The Plant Cell·Nicholas J BaltesDaniel F Voytas
Feb 28, 2014·Frontiers in Plant Science·Alexander KnollHolger Puchta
May 3, 2014·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Alexander KnollHolger Puchta
Oct 11, 2014·Nucleic Acids Research·Eva K BrinkmanBas van Steensel
Aug 22, 2015·Plant Physiology·Zhongsen LiA Mark Cigan
Nov 7, 2015·Genome Biology·Tomáš ČermákDaniel F Voytas
Dec 17, 2015·Plant Physiology·Ayako Nishizawa-YokoiSeiichi Toki
Dec 17, 2015·Plant Physiology·Masaki EndoSeiichi Toki
Jan 29, 2016·Nature Communications·Jun SongJifeng Zhang

❮ Previous
Next ❯

Citations

Oct 14, 2019·Plant Biotechnology Journal·Patrick Schindele, Holger Puchta
Nov 16, 2019·Plant Biotechnology Journal·Shaoya LiYiping Qi
Sep 1, 2020·Current Protocols in Plant Biology·Laura MerkerHolger Puchta
Dec 15, 2019·Genes·Annika Dorn, Holger Puchta
Jan 7, 2020·Critical Reviews in Biotechnology·Nikolay E ZlobinVasiliy V Taranov
Jul 16, 2020·Genes·Armin Scheben, Diego Hojsgaard
Apr 25, 2020·BMC Plant Biology·Florian HahnVladimir Nekrasov
Dec 18, 2020·Frontiers in Plant Science·Yi AnJuan Du
Jan 1, 2021·Journal of Plant Physiology·Niklas CapdevilleHolger Puchta
Dec 8, 2020·Pharmacological Research : the Official Journal of the Italian Pharmacological Society·Abhijit Dey
Jan 30, 2021·Plant Biotechnology Journal·Teng-Kuei HuangHolger Puchta
Nov 21, 2020·Frontiers in Plant Science·Anindya BandyopadhyayThomas P Brutnell
Mar 2, 2021·Transgenic Research·Teng-Kuei Huang, Holger Puchta
Mar 17, 2021·Molecular Plant Pathology·Audrey M V Ah-FongHoward S Judelson
May 30, 2021·Proceedings of the National Academy of Sciences of the United States of America·Oliver Xiaoou Dong, Pamela C Ronald
Jun 15, 2021·Journal of Genetics and Genomics = Yi Chuan Xue Bao·Qiupeng LinJin-Long Qiu
Sep 28, 2021·The CRISPR Journal·Chen-Tran HsuChoun-Sea Lin
Oct 30, 2021·Frontiers in Genome Editing·Tom LawrensonWendy Harwood
Nov 28, 2021·International Journal of Molecular Sciences·Susan Schröpfer, Henryk Flachowsky

❮ Previous
Next ❯

Related Concepts

Related Feeds

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.