This thesis explores the chemical kinetics of clustered regularly interspaced short palindromic repeats (CRISPR) enzyme systems and the design and control of microfluidic systems based on isotachophoresis (ITP). CRISPR systems constitute a recent addition to the molecular diagnostic toolbox and have sparked widespread interest over the past decade due to their easy reconfigurability and high specificity. However, the sensitivity of such systems remains limited by low enzymatic kinetic rates and background signal, especially in amplification-free formats. CRISPR kinetic rates have been widely and frequently misreported, and there has been very little quantitative estimation of the background signal. This thesis addresses these limitations through the development of quantitative kinetic models and experimental studies aimed at the rigorous determination of the sensitivity of CRISPR-based assays. ITP is a well-established electrokinetic technique that has been routinely used for sample purification and preconcentration. However, despite its versatility, its broader adoption has been limited by the complexity of assay and buffer design. This dissertation introduces computational tools, methods, and experimental demonstrations to aid the design of ITP experiments and develop complex ITP systems—including ITP-based networks for sample routing, aliquoting, and mixing.