Mycobacterium abscessus (Mabsc) infections occur in more than 5% of patients with cystic fibrosis (CF), increasing over the last decade. Antibiotic treatment for those with the infection can take many months and often are not successful at eradicating the organism, requiring long-term maintenance therapy. Currently, there is not an effective and evidence-based drug regimen for treating Mabsc infections in patients with CF. The conventional recommended treatment consists of 3-4 drugs, which together have an eradication rate as low as 20% in Mabsc subspecies abscessus. The goals of this project were: to collect experimental data to train the INDIGO-MABSC, an in silico model for predicting synergistic and antagonistic antibiotic interactions to treat M. abscessus infections; and to identify possible synergistic interactions between antibiotic combinations. I treated liquid cultures with 40 different antibiotics for 6 hours and then isolated genomic ribonucleic acid (RNA). The RNA was then sequenced to determine gene expression changes which can be used for model construction. Using broth microdilution, I set up checkerboard assays to observe the interactions between two different drugs and assess the combination’s efficiency in inhibiting the growth of Mabsc type strain ATCC19977 qualitatively by eye. I calculated the fractional inhibitory combination for each combination. Preliminary results show putative synergistic interactions between drug pairs Cefoxitin-Rifampin, Imipenem-Cefoxitin, Imipenem-Clarithromycin, and Imipenem-Linezolid. From sequencing results, we saw that Rifampin, Imipenem, Clofazimine, Rifabutin, and Cefoxitin did not induce sufficient gene changes. Future directions include additional checkerboard testing for combinations that were inconclusive due to no minimum inhibitory concentration recorded. Additionally, I will work optimize drug treatment for antibiotics that did not induce enough gene expression changes, and also work to collect RNA from cultures treated with novel drugs and antibiotic combinations that are commercially available.