Needlestick injuries (NSIs) pose significant occupational hazards for nursing students, who are at high risk due to their clinical practice. Understanding their knowledge, attitudes, and practices towards NSIs is crucial for developing effective prevention strategies. Objective: To assess the prevalence of NSIs among nursing students and evaluate their knowledge, attitude, and practice towards NSIs. Methods: This descriptive study was conducted at Sheikh Zayed Hospital Rahim Yar Khan from January 2023 to December 2023. Nursing students from the 2nd to final year were included, given their clinical exposure and high risk of NSIs. Data were collected through an online questionnaire based on previous studies, and distributed via email. Descriptive statistics were used to analyze the data, with means and standard deviations calculated for continuous variables and frequencies and percentages for categorical variables. Results: A total of 280 students participated in the survey. Sixty-seven students (23.9%) experienced NSIs, primarily during recapping (74.6%) and injections (21%). Most students (76.1%) did not report the incidents. The mean NSI knowledge score was 6.5 ± 1.3, and the mean attitude score was 27.5 ± 4.13, indicating a generally positive attitude. One hundred ninety-six students (70%) had received the Hepatitis B vaccine, 152 (54.2%) were worried about NSIs, 201 (71.7%) believed NSIs are preventable, and 150 (53.5%) felt NSIs are neglected. The mean practice score was 14.2 ± 2.1, indicating poor practice. Additionally, 140 students (50%) reported recapping needles before discarding, 98 (35%) wore gloves before injections/venipuncture, 64 (22.8%) practised one-handed recapping, 168 Conclusion: (60%) used personal protective equipment (PPE), and 56 (20%) rinsed with water and soap after an NSI. Nursing students demonstrated good knowledge and a satisfactory attitude towards NSIs; however, their practice levels were low. Enhanced training and strict adherence to safety protocols are needed to improve their practices and reduce the incidence of NSIs.