Sistersville is a city in Tyler County, West Virginia, United States, along the Ohio River. The population was 1,413 at the 2020 census. The Sistersville Ferry crosses the Ohio River to the unincorporated community of Fly in Monroe County, Ohio.

Tyler County is a county in the U.S. state of West Virginia. As of the 2020 census, the population was 8,313. Its county seat is Middlebourne. The county was founded in 1814 and is named after John Tyler, Sr., father of President John Tyler.