We report the development of a DNA hydrogel that disassembles and releases its payload in response to a target of interest. The DNA hydrogel is assembled from Y-shaped DNA motifs with polyA domains and cross-linked the small molecule cyanuric acid through hydrogen bonding. The hydrogel's structural integrity was rapidly assessed using a simple, instrumentation-free capillary migration assay that provides results within seconds. To evaluate its responsiveness to enzymatic degradation, the hydrogel was exposed to nonspecific nuclease activity using , resulting in increased mobility and decrease in fluorescence. Later, CRISPR-Cas12a was incorporated to enable programmable, target-specific hydrogel disassembly using a conserved genomic region from . Guided by crRNA sequences, the target sequences activated Cas12a to selectively degrade hydrogels. This process enabled the controlled release of various payloads, including a small-molecule drug, a fluorescent dye, a nanoparticle-based MRI contrast agent conjugated to a chemotherapeutic agent, and a model protein. To evaluate whether the hydrogel disassembly can be selectively programmed to an intended target, we tested its responsiveness against two serotypes of , , conserved genomic regions from and . To test the disassembly of this novel DNA hydrogel in the presence of a full genome, we tested the hydrogel with the genome. The target genome induced an increase in the hydrogel's mobility and loss in fluorescence with as few as 50 copies of full genome. The results demonstrate the potential of these CRISPR-responsive DNA hydrogels as intelligent platforms for target-induced imaging and therapeutic agent release, and biosensing applications.