Unlocking Nature’s Shield: The Promising Potential of CRISPRa in Amplifying Antimicrobial Peptide Expression in Common Bean (<i>Phaseolus vulgaris</i> L.)
This study proposes using the CRISPR transcriptional activation strategy to modulate the expression of genes encoding defense proteins and antimicrobial peptides (AMPs) in <i>Phaseolus vulgaris</i>. Three genes (PvD1, Pv-thionin, and Pv-lectin) were selected and targeted by the CRISPR-dCas9-TV-mediated transcriptional activation complex in the <i>P. vulgaris</i> L. hairy root. RT-qPCR investigated their activation efficiency. The eGFP-positive transgenic hairy roots exhibit enhanced expression of targeted genes compared to that of control roots. A moderate increase of 1.37-fold in PvD1 gene expression was observed in transgenic hairy roots, while 6.97-fold (Pv-lectin) and 5.70-fold (Pv-thionin) increases were observed. Importantly, no off-target effects of sgRNAs were detected, ensuring the precision and safety of the CRISPR-dCas9-TV strategy. The present article is a proof-of-concept study, and it has succeeded in demonstrating the efficiency of the CRISPR-dCas9-TV strategy in modulating the expression of target genes in <i>P. vulgaris</i>, paving the way for an alternative approach to protecting such essential crop plants.
Authors
- Cirano José Ulhôa (ORCID: https://orcid.org/0000-0002-4171-6465)
- Ângela Mehta (ORCID: https://orcid.org/0000-0002-5348-5123)
- Octávio Luiz Franco (ORCID: https://orcid.org/0000-0001-9546-0525)
- Mariana Rocha Maximiano (ORCID: https://orcid.org/0000-0002-5889-6984)
- F. J. L. Aragão (ORCID: https://orcid.org/0000-0003-0570-7673)
- Ana Cristina Miranda Brasileiro (ORCID: https://orcid.org/0000-0003-0530-062X)
- Gabriel Cidade Feitosa
- Lucas José de Sousa (ORCID: https://orcid.org/0000-0002-4747-1999)
- Osmundo Brilhante de Oliveira Neto (ORCID: https://orcid.org/0000-0002-4026-6810)
- M Lopes (ORCID: https://orcid.org/0009-0008-0416-7664)
- Bruna Medeiros Pereira
- Fabiano Touzdjian Pinheiro Kohlrausch Távora
- Brisa Ortega
- Raquel dos Santos Madeiro
Institutions
- Universidade de Brasília (BR)
- Brazilian Agricultural Research Corporation (BR)
- Universidade Católica de Brasília (BR)
- Biotech (Brazil) (BR)
- Symbiom (Czechia) (CZ)
- Centro Universitário do Distrito Federal (BR)
- Universidade Católica Dom Bosco (BR)
- Universidade Federal de Goiás (BR)
Publication Details
- Journal
- ACS Omega
- Published
- 2025-02-06
- DOI
- https://doi.org/10.1021/acsomega.4c09817
- Citations
- 2
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 1.61
Funders
- Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
- Empresa Brasileira de Pesquisa Agropecuária
- Conselho Nacional de Desenvolvimento Científico e Tecnológico
- Fundação de Apoio à Pesquisa do Distrito Federal
- Fundação de Apoio ao Desenvolvimento do Ensino, Ciência e Tecnologia do Estado de Mato Grosso do Sul