CoChat
Sign Up
Cited by 9CRISPR and Genetic EngineeringPlant tissue culture and regenerationGenetically Modified Organisms Research

DNA-free CRISPR genome editing in raspberry (Rubus idaeus) protoplast through RNP-mediated transfection

Open access
Creeth, Ryan, Thompson, Andrew, Kevei, Zoltan·2025·Frontiers in Genome Editing
DOI 10.3389/fgeed.2025.1589431PMC PMC12256493
Open this paper in CoChat — free accountOpen PDF

Abstract

Protoplast-based systems have been utilised in a wide variety of plant species to enable genome editing without chromosomal introgression of foreign DNA into plant genomes. DNA-free genome editing followed by protoplast regeneration allows elite cultivars to be edited without further genetic segregation, preserving their unique genetic composition and their regulatory status as non-transgenic. However, protoplast isolation presents a barrier to the development of advanced breeding technologies in raspberry and no protocol has been published for DNA-free genome editing in the species. Pre-assembled ribonucleoprotein complexes (RNPs) do not require cellular processing and the commercial availability of Cas9 proteins and synthetic guide RNAs has streamlined genome editing protocols. This study presents a novel high-yielding protoplast isolation protocol from raspberry stem cultures and RNP-mediated transfection of protoplast with CRISPR-Cas9. Targeted mutagenesis of the phytoene desaturase gene at two intragenic loci resulted in an editing efficiency of 19%, though estimated efficiency varied depending on the indel analysis technique. Only amplicon sequencing was sensitive enough to confirm genome editing in a low efficiency sample. To our knowledge, this study constitutes the first use of DNA-free genome editing in raspberry protoplast. This protocol provides a valuable platform for understanding gene function and facilitates the future development of precision breeding in this important soft fruit crop.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.3389/fgeed.2025.1589431

Cites

1 of 1 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Plant protoplasts allow for CRISPR screens in agriculture.

    “Murine hybridoma libraries provide monoclonal antibodies, insect cells power baculovirus expression systems, and plant protoplasts allow for CRISPR screens in agriculture [71,72,73].”

    From Genetic Insights into the Economic Toll of Cell Line Misidentification: A Comprehensive Review (2026) · cites this paper as [73]

    The Discussion reports CRISPR editing of three agriculturally relevant targets in raspberry protoplasts and says large-scale gRNA screening may be unavoidable. It does not report a CRISPR screen conducted in protoplasts, so it supports their potential for screening rather than an established screening application. (LLM, conf=0.79)

    Experimental — accuracy not yet measured

  • Murine hybridoma libraries provide monoclonal antibodies.

    “Murine hybridoma libraries provide monoclonal antibodies, insect cells power baculovirus expression systems, and plant protoplasts allow for CRISPR screens in agriculture [71,72,73].”

    From Genetic Insights into the Economic Toll of Cell Line Misidentification: A Comprehensive Review (2026) · cites this paper as [73]

  • Insect cells power baculovirus expression systems.

    “Murine hybridoma libraries provide monoclonal antibodies, insect cells power baculovirus expression systems, and plant protoplasts allow for CRISPR screens in agriculture [71,72,73].”

    From Genetic Insights into the Economic Toll of Cell Line Misidentification: A Comprehensive Review (2026) · cites this paper as [73]

    The Introduction and Discussion address CRISPR editing in plant protoplasts, not insect-cell baculovirus expression systems. Searches for “insect cells baculovirus expression systems” and “baculovirus insect cell recombinant protein expression” found no passage addressing the attributed claim. (LLM, conf=0.75)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 63

  1. Atkinson R. G. Sutherland P. W. Johnston S. L. Gunaseelan K. Hallett I. C. Mitra D. (2012). Down-regulation of POLYGALACTURONASE1 alters firmness, tensile strength and water loss in apple (Malus x domestica) fruit. BMC Plant Biol. 12, 129. 10.1186/1471-2229-12-129 22856470 PMC3509026 · DOI
  2. Barceló M. Wallin A. Medina J. J. Gil-Ariza D. J. López-Casado G. Juarez J. (2019). Isolation and culture of strawberry protoplasts and field evaluation of regenerated plants. Sci. Hortic. 256, 108552. 10.1016/j.scienta.2019.108552 · DOI
  3. Brandt K. M. Gunn H. Moretti N. Zemetra R. S. (2020). A streamlined protocol for wheat (Triticum aestivum) protoplast isolation and transformation with CRISPR-cas ribonucleoprotein complexes. Front. Plant Sci. 11, 769. 10.3389/fpls.2020.00769 32587597 PMC7298111 · DOI
  4. Brinkman E. K. Chen T. Amendola M. van Steensel B. (2014). Easy quantitative assessment of genome editing by sequence trace decomposition. Nucleic Acids Res. 42, e168. 10.1093/nar/gku936 25300484 PMC4267669 · DOI · Paper page
  5. Brockman Q. R. Scherer A. McGivney G. R. Gutierrez W. R. Rytlewski J. Sheehan A. (2023). Discrepancies in indel software resolution with somatic CRISPR/Cas9 tumorigenesis models. Sci. Rep. 13, 14798. 10.1038/s41598-023-41109-1 37684258 PMC10491828 · DOI
  6. Chan R. L. Trucco F. Otegui M. E. (2020). Why are second-generation transgenic crops not yet available in the market? J. Exp. Bot. 71, 6876– 6880. 10.1093/jxb/eraa412 32901258 · DOI
  7. Cocking E. C. (1960). A method for the isolation of plant protoplasts and vacuoles. Nature 187, 962– 963. 10.1038/187962a0 · DOI
  8. Conant D. Hsiau T. Rossi N. Oki J. Maures T. Waite K. (2022). Inference of CRISPR edits from sanger trace data. CRISPR J. 5, 123– 130. 10.1089/crispr.2021.0113 35119294 · DOI
  9. Creeth R. Thompson A. Kevei Z. (2025). DNA-Free CRISPR genome editing in raspberry (Rubus idaeus) through RNP-mediated protoplast transfection and comparison of indel analysis techniques. BioRxiv. 10.1101/2025.01.14.632935 PMC12256493 40661866 · DOI
  10. Davey M. R. Anthony P. Power J. B. Lowe K. C. (2005). Plant protoplasts: status and biotechnological perspectives. Biotechnol. Adv. 23, 131– 171. 10.1016/j.biotechadv.2004.09.008 15694124 · DOI
  11. Davik J. Røen D. Lysøe E. Buti M. Rossman S. Alsheikh M. (2022). A chromosome-level genome sequence assembly of the red raspberry (Rubus idaeus L.). PLoS One 17, e0265096. 10.1371/journal.pone.0265096 35294470 PMC8926247 · DOI
  12. Derrick S. A. Kristo A. S. Reaves S. K. Sikalidis A. K. (2021). Effects of dietary red raspberry consumption on pre-diabetes and type 2 diabetes mellitus parameters. Int. J. Environ. Res. Public Health 18, 9364. 10.3390/ijerph18179364 34501954 PMC8431376 · DOI
  13. Foster T. M. Bassil N. V. Dossett M. Leigh Worthington M. Graham J. (2019). Genetic and genomic resources for Rubus breeding: a roadmap for the future. Hortic. Res. 6, 116. 10.1038/s41438-019-0199-2 31645970 PMC6804857 · DOI
  14. Gasiunas G. Barrangou R. Horvath P. Siksnys V. (2012). Cas9–crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria. Proc. Natl. Acad. Sci. 109, E2579– E2586. 10.1073/pnas.1208507109 22949671 PMC3465414 · DOI · Paper page
  15. González M. N. Massa G. A. Andersson M. Turesson H. Olsson N. Fält A. S. (2020). Reduced enzymatic browning in potato tubers by specific editing of a polyphenol oxidase gene via ribonucleoprotein complexes delivery of the CRISPR/Cas9 system. Front. Plant Sci. 10, 497481. 10.3389/fpls.2019.01649 PMC6962139 31998338 · DOI
  16. Gou Y. J. Li Y. L. Bi P. P. Wang D. J. Ma Y. Y. Hu Y. (2020). Optimization of the protoplast transient expression system for gene functional studies in strawberry (Fragaria vesca). Plant Cell. Tissue Organ Cult. 141, 41– 53. 10.1007/s11240-020-01765-x · DOI
  17. Hsu C.-T. Lee W.-C. Cheng Y.-J. Yuan Y.-H. Wu F.-H. Lin C.-S. (2021). Genome editing and protoplast regeneration to study plant–pathogen interactions in the model plant nicotiana benthamiana. Front. Genome 2, 627803. 10.3389/fgeed.2020.627803 PMC8525392 34713245 · DOI
  18. Jeong Y. Y. Lee H.-Y. Kim S. W. Noh Y.-S. Seo P. J. (2021). Optimization of protoplast regeneration in the model plant Arabidopsis thaliana. Plant Methods 17, 21. 10.1186/s13007-021-00720-x 33622383 PMC7901198 · DOI
  19. Jia S. Wang Y. Zhang G. Yan Z. Cai Q. (2020). Strawberry FaWRKY25 transcription factor negatively regulated the resistance of strawberry fruits to Botrytis cinerea. Genes. (Basel) 12, 56– 17. 10.3390/GENES12010056 33396436 PMC7824073 · DOI
  20. Jinek M. Chylinski K. Fonfara I. Hauer M. Doudna J. A. Charpentier E. (1979)2012). A programmable dual-RNA–guided DNA endonuclease in adaptive bacterial immunity. Science 337, 816– 821. 10.1126/science.1225829 PMC6286148 22745249 · DOI · Paper page
  21. Kang H. H. Naing A. H. Kim C. K. (2020). Protoplast isolation and shoot regeneration from protoplast-derived callus of petunia hybrida cv. Mirage rose. Biol. (Basel) 9, 228. 10.3390/biology9080228 PMC7465674 32824283 · DOI
  22. Kim C. Dai W. (2022). Agrobacterium-mediated transformation of purple raspberry (Rubus occidentalis × R. idaeus) with the PtFIT (FER-like iron deficiency–induced transcription factor 1) gene. Vitro Cell. and Dev. Biol. - Plant 58, 343– 350. 10.1007/s11627-021-10228-7 · DOI
  23. Kim S. Kim D. Cho S. W. Kim J. Kim J.-S. (2014). Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins. Genome Res. 24, 1012– 1019. 10.1101/gr.171322.113 24696461 PMC4032847 · DOI · Paper page
  24. Lee S. Park S. H. Jeong Jeong Y. Kim S. Kim B. R. Ha B. K. (2024). Optimization of CRISPR/Cas9 ribonucleoprotein delivery into cabbage protoplasts for efficient DNA-free gene editing. Plant Biotechnol. Rep. 18, 415– 424. 10.1007/s11816-024-00901-9 · DOI
  25. Li R. Li Y. Zhang Y. Sheng J. Zhu H. Shen L. (2021). Transcriptome analysis reveals that SlNPR1 mediates tomato fruit resistance against Botrytis cinerea by modulating phenylpropanoid metabolism and balancing ROS homeostasis. Postharvest Biol. Technol. 172, 111382. 10.1016/J.POSTHARVBIO.2020.111382 · DOI
  26. Li R. Wang L. Li Y. Zhao R. Zhang Y. Sheng J. (2020). Knockout of SlNPR1 enhances tomato plants resistance against Botrytis cinerea by modulating ROS homeostasis and JA/ET signaling pathways. Physiol. Plant 170, 569– 579. 10.1111/PPL.13194 32840878 · DOI
  27. Lin C.-S. Hsu C.-T. Yang L.-H. Lee L.-Y. Fu J.-Y. Cheng Q.-W. (2018). Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single-cell mutation detection to mutant plant regeneration. Plant Biotechnol. J. 16, 1295– 1310. 10.1111/pbi.12870 29230929 PMC5999315 · DOI
  28. Lin C. S. Hsu C. T. Yuan Y. H. Zheng P. X. Wu F. H. Cheng Q. W. (2022). DNA-free CRISPR-Cas9 gene editing of wild tetraploid tomato Solanum peruvianum using protoplast regeneration. Plant Physiol. 188, 1917– 1930. 10.1093/plphys/kiac022 35088855 PMC8968427 · DOI
  29. Liu Y. Andersson M. Granell A. Cardi T. Hofvander P. Nicolia A. (2022). Establishment of a DNA-free genome editing and protoplast regeneration method in cultivated tomato (Solanum lycopersicum). Plant Cell. Rep. 41, 1843– 1852. 10.1007/s00299-022-02893-8 35773498 PMC9395478 · DOI
  30. López-Casado G. Sánchez-Raya C. Ric-Varas P. D. Paniagua C. Blanco-Portales R. Muñoz-Blanco J. (2023). CRISPR/Cas9 editing of the polygalacturonase FaPG1 gene improves strawberry fruit firmness. Hortic. Res. 10, uhad011. 10.1093/hr/uhad011 36960432 PMC10028403 · DOI
  31. Mathews H. Wagoner W. Cohen C. Kellogg J. Bestwick R. (1995). Efficient genetic transformation of red raspberry, Rubus ideaus L. Plant Cell. Rep. 14, 471– 476. 10.1007/BF00232777 24185514 · DOI
  32. Mezzetti B. Costantini E. Chionchetti F. Landi L. Pandolfini T. Spena A. (2004). Genetic transformation in strawberry and raspberry for improving plant productivity and quality. Acta Hortic., 107– 110. 10.17660/ActaHortic.2004.649.19 · DOI
  33. Mezzetti B. Landi L. Phan H. B. Mantovani I. Ruggieri S. Rosati P. (1999). Protoplast technology and regeneration studies for Rubus breeding. Acta Hortic., 215– 222. 10.17660/ActaHortic.1999.505.27 · DOI
  34. Moon K.-B. Park J.-S. Park S.-J. Lee H.-J. Cho H.-S. Min S.-R. (2021). A more accessible, time-saving, and efficient method for in vitro plant regeneration from potato protoplasts. Plants 10, 781. 10.3390/plants10040781 33923378 PMC8071491 · DOI
  35. Najafi S. Bertini E. D’Incà E. Fasoli M. Zenoni S. (2023). DNA-free genome editing in grapevine using CRISPR/Cas9 ribonucleoprotein complexes followed by protoplast regeneration. Hortic. Res. 10, uhac240. 10.1093/hr/uhac240 37077374 PMC10108004 · DOI
  36. Nita-Lazar M. Iwahara S. Takegawa K. Lienart Y. (2000). The active oxygen response of raspberry protoplasts to O-glycans of Fusarium sp. M7-1. J. Plant Physiol. 156, 306– 311. 10.1016/S0176-1617(00)80066-8 · DOI
  37. Park J. Choi S. Park S. Yoon J. Park A. Y. Choe S. (2019). “ DNA-free genome editing via ribonucleoprotein (RNP) delivery of CRISPR/Cas in lettuce,” in Plant genome editing with CRISPR systems. Editor Qi Y., 1917. 1st ed., 337– 354. 10.1007/978-1-4939-8991-1_25 30610648 · DOI
  38. Posé S. Paniagua C. Cifuentes M. Blanco-Portales R. Quesada M. A. Mercado J. A. (2013). Insights into the effects of polygalacturonase FaPG1 gene silencing on pectin matrix disassembly, enhanced tissue integrity, and firmness in ripe strawberry fruits. J. Exp. Bot. 64, 3803– 3815. 10.1093/JXB/ERT210 23873994 PMC3745733 · DOI
  39. Price R. J. Davik J. Fernandéz Fernandéz F. Bates H. J. Lynn S. Nellist C. F. (2023). Chromosome-scale genome sequence assemblies of the ‘Autumn Bliss’ and ‘Malling Jewel’ cultivars of the highly heterozygous red raspberry (Rubus idaeus L.) derived from long-read Oxford Nanopore sequence data. PLoS One 18, e0285756. 10.1371/journal.pone.0285756 37192177 PMC10187893 · DOI
  40. Reed K. M. Bargmann B. O. R. (2021). Protoplast regeneration and its use in new plant breeding technologies. Front. Genome 3, 734951. 10.3389/fgeed.2021.734951 PMC8525371 34713266 · DOI
  41. Rustgi S. Naveed S. Windham J. Zhang H. Demirer G. S. (2022). Plant biomacromolecule delivery methods in the 21st century. Front. Genome 4, 1011934. 10.3389/fgeed.2022.1011934 PMC9614364 36311974 · DOI
  42. Scintilla S. Salvagnin U. Giacomelli L. Zeilmaker T. Malnoy M. A. Rouppe van der Voort J. (2022). Regeneration of non-chimeric plants from DNA-free edited grapevine protoplasts. Front. Plant Sci. 13, 1078931. 10.3389/fpls.2022.1078931 36531381 PMC9752144 · DOI
  43. Scolari L. M. Hancock R. D. Graham J. Freitag S. Verrall S. Moreno-Mellado A. R. (2023). Hormone profiling in artificially induced ‘crumbly’ fruit in raspberry (Rubus idaeus L.) at two different development stages. J. Hortic. Sci. Biotechnol. 98, 384– 393. 10.1080/14620316.2022.2141140 · DOI
  44. Shan S. Soltis P. S. Soltis D. E. Yang B. (2020). Considerations in adapting CRISPR/Cas9 in nongenetic model plant systems. Appl. Plant Sci. 8, e11314. 10.1002/aps3.11314 31993256 PMC6976890 · DOI
  45. Simpson C. G. Cullen D. W. Hackett C. A. Smith K. Hallett P. D. McNicol J. (2017). Mapping and expression of genes associated with raspberry fruit ripening and softening. Theor. Appl. Genet. 130, 557– 572. 10.1007/s00122-016-2835-7 27942774 · DOI
  46. Sprink T. Wilhelm R. Hartung F. (2022). Genome editing around the globe: an update on policies and perceptions. Plant Physiol. 190, 1579– 1587. 10.1093/plphys/kiac359 35976141 PMC9614466 · DOI
  47. Sternberg S. H. Doudna J. A. (2015). Expanding the biologist’s toolkit with CRISPR-cas9. Mol. Cell. 58, 568– 574. 10.1016/j.molcel.2015.02.032 26000842 · DOI
  48. Tricoli D. M. Debernardi J. M. (2024). An efficient protoplast-based genome editing protocol for Vitis species. Hortic. Res. 11, uhad266. 10.1093/hr/uhad266 38895602 PMC11184525 · DOI
  49. Turnbull C. Lillemo M. Hvoslef-Eide T. A. K. (2021). Global regulation of genetically modified crops amid the gene edited crop boom – a review. Front. Plant Sci. 12, 630396. 10.3389/fpls.2021.630396 33719302 PMC7943453 · DOI
  50. Uusi-Mäkelä M. I. E. Barker H. R. Bäuerlein C. A. Häkkinen T. Nykter M. Rämet M. (2018). Chromatin accessibility is associated with CRISPR-Cas9 efficiency in the zebrafish (Danio rerio). PLoS One 13, e0196238. 10.1371/JOURNAL.PONE.0196238 29684067 PMC5912780 · DOI
  51. Van Campenhout C. Cabochette P. Veillard A. C. Laczik M. Zelisko-Schmidt A. Sabatel C. (2019). Guidelines for optimized gene knockout using CRISPR/Cas9. Biotechniques 66, 295– 302. 10.2144/BTN-2018-0187 31039627 · DOI
  52. Wang T. McFarlane H. E. Persson S. (2016). The impact of abiotic factors on cellulose synthesis. J. Exp. Bot. 67, 543– 552. 10.1093/jxb/erv488 26552883 · DOI
  53. Wang X. Guo R. Tu M. Wang D. Guo C. Wan R. (2017). Ectopic expression of the wild grape WRKY transcription factor VqWRKY52 in Arabidopsis thaliana enhances resistance to the biotrophic pathogen powdery mildew but not to the necrotrophic pathogen Botrytis cinerea. Front. Plant Sci. 8, 97. 10.3389/FPLS.2017.00097 28197166 PMC5281567 · DOI
  54. Wang Y. Zhang Y. Dong Y. Li D. Shi S. Li S. (2022). A highly efficient mesophyll protoplast isolation and PEG-mediated transient expression system in eggplant. Sci. Hortic. 304, 111303. 10.1016/j.scienta.2022.111303 · DOI
  55. Woo J. W. Kim J. Kwon S. Corvalán C. Cho S. W. Kim H. (2015). DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins. Nat. Biotechnol. 33, 1162– 1164. 10.1038/nbt.3389 26479191 · DOI · Paper page
  56. Woźniak-Gientka E. Agata T. Milica P. Anna B. Dennis E. Nick V. (2022). Public perception of plant gene technologies worldwide in the light of food security. Gm. Crops Food 13, 218– 241. 10.1080/21645698.2022.2111946 35996854 PMC9415543 · DOI
  57. Wright A. V. Nuñez J. K. Doudna J. A. (2016). Biology and applications of CRISPR systems: harnessing nature’s toolbox for genome engineering. Cell. 164, 29– 44. 10.1016/J.CELL.2015.12.035 26771484 · DOI
  58. Wu J. Z. Liu Q. Geng X. S. Li K. M. Luo L. J. Liu J. P. (2017). Highly efficient mesophyll protoplast isolation and PEG-mediated transient gene expression for rapid and large-scale gene characterization in cassava (Manihot esculenta Crantz). BMC Biotechnol. 17, 29– 8. 10.1186/s12896-017-0349-2 28292294 PMC5351281 · DOI
  59. Yoo S.-D. Cho Y.-H. Sheen J. (2007). Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat. Protoc. 2, 1565– 1572. 10.1038/nprot.2007.199 17585298 · DOI
  60. Yue J.-J. Yuan J.-L. Wu F.-H. Yuan Y.-H. Cheng Q.-W. Hsu C.-T. (2021). Protoplasts: from isolation to CRISPR/Cas genome editing application. Front. Genome 3, 717017. 10.3389/FGEED.2021.717017 PMC8525356 34713263 · DOI
  61. Zhan X. Lu Y. Zhu J. Botella J. R. (2021). Genome editing for plant research and crop improvement. J. Integr. Plant Biol. 63, 3– 33. 10.1111/jipb.13063 33369120 · DOI
  62. Zhang J. Yan X. Huang T. Liu H. Liu F. Yang M. (2022b). Overexpressing 4-coumaroyl-CoA ligase and stilbene synthase fusion genes in red raspberry plants leads to resveratrol accumulation and improved resistance against Botrytis cinerea. J. Plant Biochem. Biotechnol. 32, 85– 91. 10.1007/s13562-022-00784-3 · DOI
  63. Zhang Y. Cheng Y. Fang H. Roberts N. Zhang L. Vakulskas C. A. (2022a). Highly efficient genome editing in plant protoplasts by ribonucleoprotein delivery of CRISPR-cas12a nucleases. Front. Genome 4, 780238. 10.3389/fgeed.2022.780238 PMC8842731 35174354 · DOI

Cited by

Sorted by citations

Authors
Creeth, R., Thompson, A., & Kevei, Z.
Affiliations
Cranfield University (GB)
Corresponding author
Creeth, Ryan
ORCID
Venue
Frontiers in Genome Editing
Publisher
Frontiers Media SA
Published
2025-06-30
Volume
7
Pages
1589431-1589431
ISSN
2673-3439 (ISSN-L)
DOI
10.3389/fgeed.2025.1589431
PMC
PMC12256493
OpenAlex
W4411841924
Cited by
9
References
1
Citations by year
2026: 9
FWCI
2.67
Type
journal article
Language
en
Open access
Licence
Locations
  • Frontiers in Genome Editing (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • CERES (Cranfield University) (submittedVersion, cc-by)
Funders
Cranfield University
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant tissue culture and regeneration
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Genetically Modified Organisms Research
Keywords
DNA-free genome editing, CRISPR/Cas9, raspberry, protoplast isolation, phytoene desaturase, targeted mutagenesis, precision breeding, stem culture, amplicon sequencing

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review