Zahra Shahid Hussain was shot dead outside her home in Karachi by gunmen on a motorcycle.
On his Twitter feed, Imran Khan said he was holding the leader of Karachi’s dominant MQM party, Altaf Hussain, responsible for her death – a claim the MQM has strongly denied.
It comes as Karachi votes in a partial re-run of Pakistan’s general election.
Police say the motive for Zahra Shahid Hussain’s shooting is unclear.
Imran Khan said Altaf Hussain, who is in self-imposed exile in London, had “openly threatened PTI workers and leaders through public broadcasts”.
He said he was also holding the British government responsible, as he said he had warned them about Altaf Hussain.
Imran Khan tweeted his accusations from his hospital bed, where he is recovering from a back injury sustained during a fall at an election rally in Lahore.
Last week, police in London confirmed they were investigating complaints that Altaf Hussain had broken UK laws by issuing threats in a speech he made the day after the vote.
In response to accusations of electoral fraud, he is alleged to have threatened his accusers with violence.
Altaf Hussain says that his remarks were taken out of context.
He has lived in the UK since 1991, saying his life would be at risk if he returned to Pakistan.
Sunday’s electoral re-run in Karachi was ordered after Imran Khan’s party accused the MQM of widespread vote-rigging and intimidation.
The MQM – which took most of the seats in Karachi – denies any irregularities and is boycotting the vote.
Security is tight, and correspondents say tensions in Karachi are running high.
Whatever the outcome of the re-run, it will not overturn the result of last week’s vote, in which conservative leader Nawaz Sharif secured an unprecedented third term in power.
The MQM is seen as a perpetrator – as well as a victim – of violence in the city.
Since the 1980s, the MQM has won every election it has contested there.
But it also stands widely accused of ruling Karachi by fear and through vote-rigging.
The general election on May 11 marked the first transition of power from one democratically elected government since the creation of the state of Pakistan in 1947.
However, the campaign was marred by violence in which about 150 people were killed across the country.
Zahra Shahid Hussain was shot by gunmen on a motorcycle outside her home in Karachi’s upmarket Defence area, the family neighborhood of assassinated Prime Minister Benazir Bhutto.
The MQM (Muttahida Qaumi Movement) is supported mainly by Muslim Urdu-speaking people whose families moved to Sindh province at the time of the partition of India in 1947.
Voting in Karachi on May 11 was disrupted by a bomb attack outside the office of the ANP party, in which 11 people were killed and more than 40 injured.
The bombing happened in the Landhi district of Karachi, where Taliban militants are known to be active.
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