"Development of a diretional sequence and ligation-independent cloning kit".

• Grant number: 24/16813-5
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: January 01, 2026
• End date: September 30, 2026
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Rafaela Sachetto Fernandes
• Grantee: Rafaela Sachetto Fernandes
• Company: ECHO LIFE SCIENCES ASSESSORIA CONSULTORIA CIENTIFICA TECNOLOGICA LTD
• CNAE: Pesquisa e desenvolvimento experimental em ciências físicas e naturais
• Associated researchers: Rafael Victorio Carvalho Guido

Abstract

Cloning is an essential technique in molecular biology, widely used for DNA manipulation. Although traditional methods, such as the use of restriction enzymes and DNA ligases, are still widely used, they have significant limitations, including low efficiency, high cost and time-consuming processes. Modern technologies, such as In-Fusion® cloning, overcome these limitations by using homologous recombination to join DNA fragments without specific sequence requirements. However, the high cost and long import times make these kits inaccessible to most research groups in Brazil. This project proposes the development of a sequence- and ligation-independent directional cloning kit using national inputs, which is approximately 40% cheaper than the imported product. Local production of the kit will ensure rapid delivery and greater accessibility for Brazilian researchers. The technical and scientific feasibility will be assessed by standardizing an experimental protocol using national reagents, such as FastPol HF DNA polymerase, dNTPs and DH5¿ competent cells, comparing their performance to that of the In-Fusion® kit. Efforts will be directed to optimizing the cloning protocol, focusing on improving DNA polymerase activity, aiming for maximum transformation efficiency and ensuring consistent and reliable results. The market for this kit is highly promising, with the global biotechnology sector projected to grow at an average annual rate of over 19% between 2024 and 2032. The cost reduction of the kit will allow an initial 30% penetration in the national market in 3 years, with potential for export to emerging markets. As a result, this project seeks to create a competitive, affordable and high-quality national alternative, promoting technological independence and boosting the expansion of the biotechnology market in Brazil. (AU)