Williams won 23 grand slams throughout her glittering career, but is expected to retire ahead of her 41st birthday next month.
Question is, how far can the American go in what could be her last ever appearance at Flushing Meadows?
Williams has only played five singles matches since the start of the year, including a first-round loss to Emma Raducanu in Cincinnati two weeks ago.
The American tennis superstar kicks off her US Open campaign against Montenegro's Danka Kovinic.
Williams has struggled with a knee injury ahead of the tournament, though she heads into her clash against the world number 80 having fared well in practise.
Elsewhere, Andy Murray will be one of the first players on court as he kicks off his latest US Open campaign against Argentina's Francisco Cerundolo.
The former world number one underwent a number of medial tests before the event as he tried to resolve recent cramp issues.
Results failed to flag up any issues, despite the fact Murray has struggled with cramp in three different matches during the Cincinnati Open last week.
Nevertheless, the 2012 US Open champion will be determined to overturn a poor run of form which has seen him lose four of his last five games.
Current world number one Daniil Medvedev will also be in action on the opening day. The Russian will take on Stefan Kozlov as he looks to close out the 2022 season in style.
The 26-year-old is the defending champion at Flushing Meadows, having pipped Novak Djokovic to the US Open crown in 2021.
Match to watch - Nick Kyrgios vs Thanasi Kokkinakis
Australian friends and teammates will meet head-to-head in the first round of the US Open as Nick Kyrgios squares off with Thanasi Kokkinakis.
A sold-out Arthur Ashe stadium will expect a spectacle as the in-form Kyrgios plots another successful grand slam run.
Kokkinakis is also bracing himself for what he described as a 'circus', saying in his pre-tournament press conference: "He loves to kind of live in the chaos a bit when it comes to matches. He turns it into a bit of a circus around him and he's just so comfortable with that.
"He's done it from such a young age. Even 12 and under, 13 and under nationals - I've known him for a while. But he's got the game, obviously, to back it up. It's not just that, so it will be awkward."
Potential upset - Tatjana Maria vs Maria Sakkari
The battle of the Maria's could be one of the most scintillating clashes of the opening day.
While Sakkari enters the match as the world number three and as the bookies favourite, though she has been engulfed by tough times since Indian Wells.
The Greek player has won 12 matches from 12 tournaments, with Germany's Tatjana Maria poised to take full advantage of her opponent's poor form.
Maria reached the Wimbledon semi-final last month, though she has struggled to replicate her winning formula since.