The National Olympic Committee of Lithuania (Lithuanian: Lietuvos tautinis olimpinis komitetas, LTOK) fielded a team of 67 athletes, 48 men and 19 women, across 15 sports at the Games.

The Lithuanian squad featured three Olympic medalists from London 2012, namely light welterweight boxer Evaldas Petrauskas, breaststroke swimmer and world record holder Rūta Meilutytė, and defending champion Laura Asadauskaitė in the women's modern pentathlon.

Track cyclist Simona Krupeckaitė topped the list of experienced Lithuanian athletes to make her fourth Olympic appearance, while Laser Radial sailor and Beijing 2008 silver medalist Gintarė Scheidt was selected to lead the Lithuanian delegation as the flag bearer in the opening ceremony at her third Games, a historic first by a female in the nation's Olympic history.

[5] Three of these medals were awarded to the Lithuanian team in weightlifting and rowing for the first time in 16 years, including a silver-medal feat from Griskonis and his new partner Saulius Ritter in the men's double sculls.

Meanwhile, Aurimas Lankas and Edvinas Ramanauskas continued to add canoeing medals for Lithuania with a bronze in the men's K-2 200 m, following Jevgenijus Šuklinas' runner-up finish at London 2012.

[15] One additional spot was awarded to the Lithuanian cyclist in the women's road race by virtue of her top 100 individual placement in the 2016 UCI World Rankings.

Santa Pakenytė earned a continental quota spot from the European region as the highest-ranked Lithuanian judoka in the IJF World Ranking List of 30 May 2016.

[29] Lithuania has qualified one wrestler for the men's Greco-Roman 66 kg into the Olympic competition as a result of his wrestle-off triumph at the initial meet of the World Qualification Tournament in Ulaanbaatar.