The first trial of VAR at a WSL stadium will take place in Chelsea's friendly against AS Roma at Kingsmeadow on Sunday.
It was first used at an English domestic women's game during the 2023 FA Cup final when Chelsea beat Manchester United 1-0 at Wembley.
The technology trial will be overseen by refereeing body PGMOL and the FA.
"This is the next step in testing VAR technology in the women's game," said PGMOL's Bibi Steinhaus-Webb.
"The preparation for the introduction of VAR technology has been happening for a sustained period of time now, and officials continually take part in both theory and on-field sessions, including behind-closed-doors practice matches."
Steinhaus-Webb, an elite-level German former referee who now serves as PGMOL's women's select group director, added: "If the support of VAR technology is needed in this weekend's match between Chelsea and AS Roma, it will be there and the teamwork between the on-field match officials and VAR team will be crucial to achieving the correct outcome."
Abi Byrne is due to be the matchday referee. Emily Heaslip and Chloe-Ann Small will operate as video assistant referee (VAR) and assistant VAR respectively from the PGMOL VAR facility at Stockley Park near London when the match kicks off at 8pm BST.
Heaslip was the referee when VAR was deployed in the Women's FA Cup final in May.
VAR was used on that occasion to confirm the ruling out of a goal by United's Leah Galton and to clarify a foul should result in a free-kick rather than a penalty when Nikita Parris was taken down by Niamh Charles.
Hannah Buckley, the Football Association's senior facilities and projects manager, said the VAR trial would be "used as part of our ongoing work to prepare match officials and clubs for its potential use should it be considered for wider implementation in the future".