The Czech Republic has been very actively involved in UNESCO's activities from the very beginning. It is a member of most of the international conventions negotiated under the UNESCO regime and a number of international and intergovernmental programmes established by the UNESCO General Conference. It is also active in a number of intergovernmental bodies of these conventions and programmes (e.g. the Committee for the Protection of Cultural Property in Armed Conflict, the Headquarters Committee, the Executive Committee for the International Campaign for the Establishment of the Museum of Nubia in Aswan and the National Museum of Egyptian Civilisation in Cairo, the Intergovernmental Council of the Management of Social Transformations Programme - MOST). The President of the 30th session of the General Conference from 1999 to 2001 was Jaroslava Moserová, the then President of the Czech Commission for UNESCO. The Czech Republic was a member of the Executive Board from 1995 to 1999, 2003 to 2007 and 2011 to 2015. The Czech Republic is currently being considered for reappointment to the Executive Board.

The Czech Republic participates in regional consultations and discussions in the framework of the implementation of Sustainable Development Goal 4 of the 2030 Agenda, i.e. ensuring inclusive and equitable quality education and promoting lifelong learning opportunities for all. The Chairman of the Czech Commission for UNESCO, Prof. Stanislav Štech, PhD, CSc., is the UNESCO Constituency II representative on the Steering Committee for the implementation of the SDGs in the field of education.  As early as 1966, Czechia joined the UNESCO Associated Schools Network project, which now includes 52 primary and elementary art schools, grammar schools, secondary schools, vocational secondary schools and higher vocational schools in the Czech Republic. These schools actively promote UNESCO's ideas, which they incorporate into their curricula, and guide their pupils towards the four basic pillars of education: learning to know, learning to do, learning to live together.

There are currently six Biosphere Reserves in the Czech Republic: Třeboňsko (1977), Křivoklátsko (1977), Šumava (1990), the bilateral Czech-Polish Krkonoše/Karkonosze (1992), Bílé Karpaty (1996) and Dolní Morava (2003), which are areas not only of valuable natural ecosystems but also promote harmonious relations between the environment and its inhabitants. Biosphere reserves are designated under one of UNESCO's oldest programmes, Man and the Biosphere. There is also one global geopark in the Czech Republic - Bohemian Paradise (2005).

The Czech Republic currently has 16 monuments (15 cultural and 1 natural) on the World Heritage List: the Historical Centre of Prague (1992) and Průhonice Park (2010), the Historical Centre of Český Krumlov (1992), the Historical Centre of Telč (1992), the Pilgrimage Church of St. Barbora, Cathedral of the Assumption of the Virgin Mary in Sedlec (1995), Lednice-Valtice Cultural Landscape (1996), Gardens and Chateau in Kroměříž (1998), Historical Village of Holašovice (1998), Litomyšl Chateau and Chateau Complex (1999), Holy Trinity Column in Olomouc (2000), Villa Tugendhat in Brno (2001), Jewish Quarter and St. Prokop in Třebíč (2003), Mining Region Erzgebirge/Erzgebirge (2019, together with Germany), Landscape for Breeding and Training of Ceremonial Carriage Horses in Kladruby nad Labem (2019), Famous Spa Towns of Europe (2021, together with Germany, Austria, Belgium, France, Italy and the United Kingdom) - the Czech Republic is represented by the spa towns of Františkovy Lázně, Karlovy Vary and Mariánské Lázně. The natural monument on the list is the beech forests of the Jizera Mountains (2021), which belong to the Ancient and Primeval Beech Forests and Forests of the Carpathian Mountains and other areas of Europe (a transnational serie of natural heritage sites). The Czech Republic has also submitted a nomination for Žatec and the landscape of Žatec hops, which is in the process of evaluation.

The Czech Republic has 7 items on the Representative List of the Intangible Cultural Heritage of Humanity: The Slovácko verbuňk (2005), The village Shrovetide processions in the Hlinecko area (2010), Falconry - living human heritage (2010), The Ride of the Kings as a part of the Pentecost traditions the South-East Czechia (2011), Puppetry in Slovakia and the Czech Republic (2016), Blueprint - resist hand printing and indigo dyeing in Europe  (2018), and Handmade ornaments from blown glass beads for Christmas trees (2020).

The Czech Republic also has eight documentary heritage sites inscribed on the international Memory of the World Register: Collection of Medieval Manuscripts from the Czech Reformation (2007), Collection of Russian, Ukrainian and Belarusian Emigrant Periodicals from 1918 to 1945 (2007), Prague Collection of 526 Graphic University Theses from 1637 to 1754 (2011), "Libri prohibiti" - a collection of Czech and Slovak samizdat periodicals from 1948 to 1989 (2013), Émile Reynaud's Light Pantomime (original projection strip) (2015, together with France), Leoš Janáček Archive (2017), Kynžvart daguerreotype (2017), and Camocio's maps of the Great Siege of Malta from 1565 (2017, together with Malta).