Genetic manipulation in bacteria, particularly in industrially relevant Streptomyces strains, is often hindered by low DNA transfer efficiency. This study investigates the role of cell envelope components, particularly teichoic acid (TA), in enhancing conjugation efficiency. Using CRISPR-Cpf1 system, we systematically disrupted 26 cell envelope-related genes in Streptomyces coelicolor A3(2), revealing that TA biosynthesis significantly influences DNA uptake. Deletion of SCO1526 (phosphatidylinositol mannoside acyltransferase) and SCO4847 (d-alanyl-d-alanine carboxypeptidase) markedly increased conjugation efficiency, while heterologous expression of TA biosynthetic genes in industrial strains S. hygroscopicus, S. avermitilis and S. venezuelae resulted in a 1300-fold, 4.9-fold and 4.9-fold enhancement, respectively. Strain-specific differences in TA impact probably linked to variations in cell wall structure and TA synthesis capacity. These findings highlight the critical role of TA in bacterial conjugation and offer a robust strategy for improving genetic manipulation in industrially important Streptomyces strains. This work advances our understanding of bacterial genetic tractability and provides a foundation for harnessing the biosynthetic potential of traditionally hard-to-manipulate bacteria.