The advancement in medical science has created health care environments that require nursing professionals who posses specialized clinical knowledge and skills to provide care and deal with critically and acutely ill children. This study explored gaps between knowledge and practice as perceived by the registered nurses of pediatric units by further recommending the changes suggested by them. A descriptive exploratory Study Design under the quantitative research methodology was utilized using universal sampling of all pediatric nurses working at a tertiary care hospital in Karachi, Pakistan. The gaps between knowledge and practice, as perceived by the participants, were categorized into five major categories: (1) medication (34%), (2) skills (28.3%), (3) knowledge (13.36%), (4) handling of code blue and intubations (12.6%), and (5) operating medical devices (11.58%). As a result, anxiety and incompetency were notably seen in the participants which had great amount of impact on the level of care provided to the Patients. The implications of the findings for quality Patient care were also analyzed.