Background: Problem-based learning (PBL) is now a well established method in medical education in many medical schools in the world. Assessing knowledge and perception of the medical staff towards PBL is important, particularly in medical colleges which have not yet introduced this method. Objective: Assessing knowledge, attitude, and practice of the medical staff at Faculty of Medicine (Assiut), Al-Azhar University towards problem-based learning approach. Methods: A cross-sectional study was conducted at the previously mentioned setting during the study year 2015/16 where the PBL teaching method was still un-applied. All medical staff (n= 178) of the college was invited to participate in this study. Relevant data about staff characteristics as well as their knowledge, attitude and practice towards PBL were collected through a predesigned structured questionnaire.   Results: The overall response rate of the medical staff was 74.7% (133 out of 178). The proportion of staff having good knowledge about PBL was (69.2%) with the highest percentage of good knowledge  was insignificantly observed among staff aged  50+ years(83.3%), male staff (70.2%) and staff of academic departments (74.2%), while professors had significantly the highest percentage(85.3%) of good knowledge. There was also an overall favorable attitude (88%) among the staff towards PBL. The mean score of favorable attitude was insignificantly higher among staff aged 50+ years, female staff and staff of academic departments, while assistant professors had significantly the higher mean score of favorable attitude than other staff grades. A small proportion of the studied staff (7.5%) has practiced PBL before. Conclusion: A reasonable proportion of the medical staff was found to have good knowledge and favorable attitude towards PBL, while previous practice of PBL was low among the staff.  Preparing educational courses about PBL as newly suggested teaching methods is recommended for all staff in terms of its definition, advantages and outcome for the college development and quality of the teaching process.