The Agave genus is recognized for its diversity, economic importance, and adaptability to arid and semi-desert climates. Among its species, Agave sisalana stands out for the production of sisal, a resilient natural fiber with various industrial applications and potential to be used as a raw material in biofuel production, due to the accumulation of fermentable sugars in its biomass. Although this species presents significant agronomic potential, challenges in conventional breeding hinder its cultivation. A viable alternative for genetic improvement is the development of transgenic plants that incorporate desirable agronomic traits through the controlled insertion of genes of interest into their genome. The objective of study was to develop a protocol for obtaining transgenic plants of Agave sisalana 'RLV19,' a species widely cultivated in the semi-arid region of Bahia-BA, Brazil, using the cp4-epsps gene aimed at constitutive gene expression. Two transgenic plants of Agave sisalana 'RLV19' were regenerated via organogenesis. PCR analyses and CP4-EPSP protein expression by RT-PCR confirmed the presence and expression of the transgene in these plants. This is the first report of A. sisalana transgenic plants expressing the cp4-epsps gene, and to the best of our knowledge, there have been no prior reports on protocols for the production of transgenic plants of this species.