Herein, we present the comprehensive in vitro and in vivo evaluation of this optimized microneedle patch, demonstrating its (i) sufficient mechanical strength for reliable skin penetration; (ii) rapid skin barrier recovery and excellent biosafety; (iii) tunable, enzyme-free glucose-responsive insulin-release kinetics in vitro; and (iv) superior efficacy in maintaining stable and safe glycemic control in vivo in a type 1 diabetic mouse model, critically mitigating the risk of the hypoglycemic nadir common with SC injections. The gene discussed is INS; the disease is type 1 diabetes mellitus.