Technical Abstract: Storing potato (Solanum tuberosum) tubers at cold temperatures prevents sprouting and minimizes losses due to disease. Unfortunately, cold storage triggers an accumulation of reducing sugars, a phenomenon referred to as cold-induced sweetening (CIS). High-temperature processing of potato tubers with elevated amounts of reducing sugars results in dark-colored, bitter-tasting products that are unacceptable to consumers. These products also have elevated amounts of acrylamide, and acrylamide has become a major food safety concern. We set out to control both problems by silencing the potato gene encoding vacuolar invertase (VInv). Vacuolar invertase hydrolyzes sucrose to form the reducing sugars glucose and fructose and this activity is the final step in a metabolic pathway in which starch breakdown leads to reducing sugar accumulation during CIS. Potato chips made from tubers derived from RNAi-based VInv gene silencing lines had acceptable, light color after cold storage for up to six months. Chip color was directly correlated with the amount of VInv transcript among RNAi lines. In addition, acrylamide content was reduced up to 15-fold in chips processed from VInv gene silencing lines compared to controls. Plant growth characteristics, yield, and specific gravity for most RNAi lines were not different from untransformed controls. The data demonstrate that the VInv gene plays a critical role in CIS. Suppressing this gene can be employed to effectively solve both processing quality and acrylamide problems in potato. The relationship between VInv gene expression and chip color can be used to establish a benchmark for use in developing new potato varieties that produce light-colored chips with low acrylamide after low temperature storage.