In footage posted to Ms Greene’s Twitter account on Friday, she beckons two masked people to pass through the “checkpoint”, consisting of a rope and stand placed in the middle of the Rotunda.
“That’s the House side, Nancy Pelosi will have you arrested,” she whispers while smirking and shooting a glance at the camera.
“This is the Senate side, come on.”
Soon afterwards the rope and stands are taken away by security.
“Democrats really hate walls. Especially my COVID border checkpoint,” Ms Greene writes on Twitter.
The reintroduction of compulsory mask use in the House has provoked days of performative outrage by Republicans.
On Thursday, 40 maskless Republican lawmakers including Ms Greene and Lauren Boebert burst into the Senate chamber to protest the regulations.
Ms Boebert is said to have thrown a face mask back at a congressional staffer after being asked to wear one and Ms Greene previously shredded a fine for refusing to wear a face covering.
The protests prompted the Capitol police chief Thomas Manger to order officers to report any members of Congress who refuse to wear a mask, warning they would be subject to arrest for unlawful entry under DC Code 22-3302.
Meanwhile, House Republican representatives Thomas Massie and Nancy Mace dared Pelosi and police to come at them.
The Congress mask mandate came as cases of the Delta variant are rising in all 50 states, and the Centers for Disease and Control Prevention advised vaccinated Americans living in areas of high or substantial transmission to wear masks when indoors.
On Thursday the World Health Organisation confirmed the US had the most reported new cases of any country in the world last week, with just over 500,000 people becoming infected with the virus.