The competition gets underway with hosts Cameroon taking on Burkina Faso at the Olembe Stadium in Yaounde.
The organisers, the Confederation of African Football (CAF), had previously used VAR from the quarter-finals stage of the 2019 edition in Egypt.
However, CAF has now advanced with a full use of the innovation which gained worldwide recognition by 2018.
It is expected to help improve the image of refereeing and also produce world-class match officials in Africa.
Sixty-three match officials, including women referees Salima Mukasanga (Rwanda), Carine Atemzabong (Cameroon), Fatiha Jermoumi (Morocco) and Bouchra Karboubi (Morocco), have been listed for the matches.
The list consists of 24 referees, 31 assistant referees and eight VARs, all coming from 36 countries.
The roster includes; two referees from the CONCACAF as part of an inter-confederation skills exchange programme.
All 52 AFCON matches in Cameroon to use VAR- CAF