Resistance of mosquitoes to insecticides is a growing concern in India. Since only a few insecticides are used for public health and limited development of new molecules is expected in the next decade, maintaining the efficacy of control programs mostly relies on resistance management strategies. Developing such strategies requires a deep understanding of factors influencing resistance together with characterizing the mechanisms involved. Among factors likely to influence insecticide resistance in mosquitoes, agriculture and urbanization have been implicated but rarely studied in detail. In the present study, we evaluate the permethrin resistance and cross-resistance pattern of several insecticides in Culex quinquefasciatus mosquitoes. After 10 generation of selection with permethrin, the LC50 value for both larvae and adult Cx. quinquefasciatus was increased by 17.3- and 17.1-folds compared with susceptible strain. Detoxification enzyme profiles and native PAGE electrophoresis of esterase isoenzyme further revealed that esterase and CytP450 may be involved in permethrin resistance (PerRes) strain compared with susceptible strain. In addition to cross-resistance, study revealed that high resistance to cypermethrin (RR = 6.3, 8.8-folds). This study provided important information for understanding permethrin resistance and facilitating a better strategy for the management of resistance. These studies conclude that a strong foundation for further study of permethrin resistance mechanisms observed in Cx. quinquefasciatus mosquitoes.