Knox was sentenced last year to 26 years in prison for the death of Meredith Kercher at a villa the two shared in Perugia, the central Italian town where both were students.
Knox has vehemently proclaimed her innocence and her family has continued to fight the conviction.
There was nearly two hours of legal wrangling between attorneys before the judge decided to allow inmate Mario Alessi to testify. Alessi is serving a 30-year sentence for kidnapping and killing a 18-month old boy.
Alessi testified that Rudy Guede, who has also been convicted of being involved in Kercher's murder and is serving a 16-year sentence, told him that Knox was not involved in the killing.
Gallery: Amanda Knox's childhoodAccording to Alessi, Guede said that he and an unnamed friend visited Kercher to try to get her to participate in sexual acts. When Kercher denied them, Guede's friend forced himself on her, and Guede also participated, Alessi said. At one point, the friend pulled out a knife, and then Guede, who was holding Kercher at the time, noticed she had started bleeding, Alessi testified.
Alessi told the court that Guede tried to stop the bleeding, but that his friend urged him to leave. Alessi said Guede remained by himself with Kercher for a while, but eventually left her injured.
Prosecutors questioned the validity of Alessi's testimony.
Through his lawyers, Guede has denied this version of events.
Another witness Saturday was Marco Castelluccio, a mafia member who testified from behind a screen because he is a protected witness.
Castelluccio said that he also heard from Guede that Knox had nothing to do with the crime.
Another inmate expected to testify is Luciano Aviello, who is serving a 16-year prison term for mafia-related crimes. Aviello is expected to say that it was his brother and Guede who killed Kercher.
Kercher, 21, was found dead on November 2, 2007, semi-naked with her throat slashed. Knox and her former Italian boyfriend, Raffaele Sollecito, were both found guilty of the murder. Knox was sentenced in December 2009.
Guede, a drifter originally from the Ivory Coast, was tried separately.
As part of her appeal, forensic experts are retesting evidence that was used to convict her.
That evidence includes a knife found in Sollecito's apartment with Knox's DNA on the handle and what Perugia prosecutors say is Kercher's DNA in a tiny groove on the blade.
The prosecution contends that the knife was used to stab Kercher in the neck and that it had been cleaned. The DNA matter attributed to Kercher consists of flesh, not blood, they say.
The sample, however, was so small that forensic scientists investigating Kercher's murder were not able to double test it in accordance with international forensic science norms, which Knox's legal team says raises doubts about its validity.
The second piece of evidence the forensic experts are testing is the tiny metal clasp from Kercher's bra, which was cut from her body after her slaying. Forensic scientists in the investigatory phase determined that Sollecito's DNA is present on the metal clasp.
The clasp was identified on an investigatory video the same day Kercher's body was found. But it was not collected until nearly six weeks later, giving the defense cause to question whether the sample may have been contaminated.
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