Abstract: Detecting small molecules and other biologically important targets, such as signaling molecules, drugs, toxins, and oligonucleotides, presents unique challenges in complex native environments and at low target concentrations. We developed field-effect transistors (FETs) coupled with nucleic-acid receptors (i.e., aptamers), for sensing in situ. Rare aptamer sequences are identified via solution-phase SELEX, thereby circumventing target tethering and epitope masking. Rigorous counter-SELEX against similarly structured metabolites and interferents yields aptamers with high target selectivity. Oligonucleotide libraries are designed for stem closure and adaptive loop-binding upon target recognition. Target-induced conformational rearrangements of stem-loop aptamers are transduced into conductance changes at nanometer-thin In2O3 FET semiconductor surfaces. Portions of the conformational changes in highly negatively charged nucleic acid backbones occur within the Debye length (<1 nm in physiological fluids) to enable direct target quantification over 5-6 orders of magnitude and at concentrations well below aptamer-target dissociation constants. We have demonstrated selective sensing of small-molecule neurotransmitters (e.g., serotonin, dopamine) in brain tissue and nutrients (e.g., glucose, phenylalanine) in blood. Via hybridization, we detect and differentiate single-nucleotide polymorphisms sans amplification. Paths to temporally resolved in vivo sensing and other key applications will be illustrated. Metal-oxide thin-film FETs fabricated via sol-gel processing, chemical-vapor deposition, standard and novel low-cost chemical patterning methods, and on flexible substrates enable multiplexed sensing with wide accessibility and applicability.