ArticlePlant biotechnology journal2017
CRISPR-Cas9-mediated efficient directed mutagenesis and RAD51-dependent and RAD51-independent gene targeting in the moss Physcomitrella patens.
Article in Plant biotechnology journal, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 63 papers, 1 of them a synthesis that pooled it.
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Who cites it
63 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Which Factors Affect the Occurrence of Off-Target Effects Caused by the Use of CRISPR/Cas: A Systematic Review in Plants.Frontiers in plant science · 2020Pooled it
- Recombinant Protein Drugs: A 2025 Update.BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy · 2026Review
- The Importance of thePlants (Basel, Switzerland) · 2026Review
- tRNA-based polycistronic CRISPR/Cas9 system boosts efficiency of multi-gene deletion in the moss Physcomitrella.Plant methods · 2026Article
- Essential developmental processes in Physcomitrium patens require distinct levels of total activity provided by functionally redundant PpROP GTPases.The New phytologist · 2025Article
- Complex sphingolipid metabolism impacts cell division and plasmodesmal development in the moss Physcomitrium patens.Plant physiology · 2025Article
- CRISPR/Cas9-Induced Targeted Mutagenesis of the MossBio-protocol · 2025Article
- Moss BRCA2 lacking the canonical DNA-binding domain promotes homologous recombination and binds to DNA.Nucleic acids research · 2025Article
- The Extended Synaptotagmins ofPlants (Basel, Switzerland) · 2025Review
- Class II kinesin-12 facilitates cell plate formation by transporting cell plate materials in the phragmoplast.Nature plants · 2025Article
- CRISPR/Cas9 targeted genetic screening in Physcomitrella identifies novel cell division genes.Frontiers in plant science · 2025Article
- The transcription factor PpRKD evokes female developmental fate in the sexual reproductive organs of Physcomitrium patens.The New phytologist · 2025Article
- Cycling DOF factor mediated seasonal regulation of sexual reproduction and cold response is not conserved inPlant direct · 2024Article
- The SpRY Cas9 variant release the PAM sequence constraint for genome editing in the model plant Physcomitrium patens.Transgenic research · 2024Article
- Development of PEG-mediated genetic transformation and gene editing system of Bryum argenteum as an abiotic stress tolerance model plant.Plant cell reports · 2024Article
- Two subtypes of GTPase-activating proteins coordinate tip growth and cell size regulation in Physcomitrium patens.Nature communications · 2023Article
- Improved prime editing allows for routine predictable gene editing in Physcomitrium patens.Journal of experimental botany · 2023Article
- Armadillo repeat-containing kinesin represents the versatile plus-end-directed transporter in Physcomitrella.Nature plants · 2023Article
- CRISPR-Cas9 Genome Editing in the Moss Physcomitrium (Formerly Physcomitrella) patens.Current protocols · 2023Article
- Control of Plant Cell Growth and Proliferation by MO25A, a Conserved Major Component of the Mammalian Sterile 20-Like Kinase Pathway.Plant & cell physiology · 2023Article
3 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
9 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The ability to address the CRISPR-Cas9 nuclease complex to any target DNA using customizable single-guide RNAs has now permitted genome engineering in many species. Here, we report its first successful use in a nonvascular plant, the moss Physcomitrella patens. Single-guide RNAs (sgRNAs) were designed to target an endogenous reporter gene, PpAPT, whose inactivation confers resistance to 2-fluoroadenine. Transformation of moss protoplasts with these sgRNAs and the Cas9 coding sequence from Streptococcus pyogenes triggered mutagenesis at the PpAPT target in about 2% of the regenerated plants. Mainly, deletions were observed, most of them resulting from alternative end-joining (alt-EJ)-driven repair. We further demonstrate that, in the presence of a donor DNA sharing sequence homology with the PpAPT gene, most transgene integration events occur by homology-driven repair (HDR) at the target locus but also that Cas9-induced double-strand breaks are repaired with almost equal frequencies by mutagenic illegitimate recombination. Finally, we establish that a significant fraction of HDR-mediated gene targeting events (30%) is still possible in the absence of PpRAD51 protein, indicating that CRISPR-induced HDR is only partially mediated by the classical homologous recombination pathway.
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