Although blood-based liquid biopsy is a promising noninvasive technique to acquire a comprehensive molecular tumor profile by detecting cancer-specific biomarkers (e.g. DNA, RNA, and proteins), there has been limited progress for brain tumor application partially because the low permeability of the blood-brain barrier (BBB) hinders the release of tumor biomarkers. We previously demonstrated focused ultrasound-enabled liquid biopsy (FUS-LBx) that uses FUS to increase BBB permeability in murine glioblastoma models and thus enhance the release of tumor-specific biomarkers into the bloodstream. The objective of this study was to evaluate the feasibility and safety of FUS-LBx in the normal brain tissue of a porcine model. Increased BBB permeability was confirmed by the significant increase (p = 0.0053) in K trans (the transfer coefficient from blood to brain extravascular extracellular space) when comparing the FUS-sonicated brain area with the contralateral non-sonicated area. Meanwhile, there was a significant increase in the blood concentrations of glial fibrillary acidic protein (GFAP, p = 0.0074) and myelin basic protein (MBP, p = 0.0039) after FUS sonication as compared with before FUS. There was no detectable tissue damage by t 2 *-weighted MRi and histological analysis. findings from this study suggest that FUS-LBx is a promising technique for noninvasive and localized diagnosis of the molecular profiles of brain diseases with the potential to translate to the clinic. Tissue biopsy has been used to characterize and track the tumor molecular landscape. However, tissue biopsy for brain tumor diagnosis requires invasive surgical procedures, which carry a 5-7% risk of major morbidity 1. It may not be possible at all to perform this procedure on medically inoperable patients or patients with tumors in surgically inaccessible locations. Repeated tissue biopsies to assess treatment response and cancer recurrence are often not feasible given the increased risk for complications and morbidity. These challenges limit the timely diagnosis and selection of treatment options, hinder a better understanding of the disease, and impair the development of effective treatment approaches. Liquid biopsy (LBx), which refers to the detection of tumor-derived components in body fluids (e.g., blood, urine, saliva, ascitic fluid, cerebrospinal fluid, etc.), has been gaining enormous attention in both medical research and clinical applications 2,3. Various substances from liquid biopsies have been found to be closely related to the stage of a tumor and might serve as biomarkers for cancer diagnosis and prognosis, such as circulating tumor cells, circulating tumor DNAs, RNAs, extracellular vesicles, and a series of cancer-related proteins 4. Blood-based LBx enables physicians to noninvasively interrogate the dynamic evolution of a tumor and monitor a patient's response to therapies through a simple blood test. Blood-based LBx-guided personalized therapy has already entered clinical practice in the management of several cancers. An import...