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1 year oldTamara Lich and Chris Barber each face counts of mischief and obstructing police.
The two were part of a group that led a convoy of lorries to Ottawa to protest Covid-19 measures and Prime Minister Justin Trudeau's government.
Experts say the outcome of the trial could reverberate beyond the courts.
The main charge that Mr Lich and Mr Barber will have to defend is that of mischief, defined under Canadian law as the wilful destruction, damaging, obstruction, or interference of property.
Joao Velloso, a law professor at the University of Ottawa, said legally, the case overall is quite ordinary.
But he anticipates an abnormal level of attention given its tie to the Freedom Convoy protests and the political tensions around issues like vaccine mandates and freedom of assembly.
The outcome of the case will be viewed differently depending on where people stand on these issues and their overall trust in the justice system, he said.
"In that sense, (the case) is bigger than the law itself."
The February 2022 protests were initially sparked by a federal vaccine mandate for lorry drivers crossing the US- Canada border.
Convoys of some 400 heavy trucks and other vehicles descended on Ottawa, Canada's capital, and blockaded city streets around parliament for three weeks.
City officials deemed it an "occupation".
Separate protests also blocked a key US-Canada border crossing near Detroit, angering the White House and disrupting the flow of goods. Other border points were blocked in Alberta and Manitoba.
The protests received international attention and inspired similar copycat demonstrations abroad.
They came to an end after Mr Trudeau invoked the Emergencies Act - the first time the Canadian law has been used - which allowed police to clear the streets and the government to impose bans on public assembly and freeze the bank accounts of protesters.
Ms Lich and Mr Barber were arrested shortly afterwards.
Paul Daly, Chair in Administrative Law and Governance at the University of Ottawa, said a key question will be to what extent the pair were responsible for the disruption experienced by Ottawa residents.
Many complained of excessive noise, daily disruptions and instances of harassments during the protest.
The trial "pits the free speech and free association rights of convoy organisers against the public interest in keeping city streets liveable for residents," Mr Daly said.
Lawrence Greenspon, Ms Lich's lawyer, said in a statement to the BBC that "we do not expect this to be the trial of the Freedom Convoy".
"The central issue will be whether the actions of two of the organisers of a peaceful protest should warrant criminal sanction."
Still, during a recent book event, Ms Lich said she believes the outcome of the trial will set a precedent for other protestors also awaiting trial.
"That's why I'm not going down without a fight and we will come out swinging," she said in July.
Crown prosecutors have argued that, if Ms Lich is found guilty, she could face a sentence of up to 10 years in prison.
But during a July bail hearing, an Ontario court judge said that due to this "very unusual case, there is significant uncertainty about the degree to which she will be held culpable for the assortment of alleged bad acts committed over many weeks by various persons in a crowd of thousands".
Both Ms Lich and Ms Barber, along with other named organisers, are facing a separate C$300m ($222m; £175m) civil suit brought against them by Ottawa residents over the disruptions in the city.
The trial of Ms Lich and Mr Barber is set for 13 days, followed by another six days in October.
Pat King, another organiser of the protests, is facing a separate trial in November.
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