The persecution of Christians in the New Testament is an important part of the Early Christian narrative which depicts the early church as being persecuted for their heterodox beliefs by a Jewish establishment in the Roman province of Judea. The New Testament, especially the Gospel of John, has traditionally been interpreted as relating Christian accounts of the Pharisee rejection of Jesus and accusations of the Pharisee responsibility for his crucifixion. The Acts of the Apostles depicts instances of early Christian persecution by the Sanhedrin, the Jewish religious court.

Walter Laqueur argues that hostility between Christians and Jews grew over the generations. By the 4th century, John Chrysostom was arguing that the Pharisees alone, not the Romans, were responsible for the murder of Christ. However, according to Laqueur: "Absolving Pilate from guilt may have been connected with the missionary activities of early Christianity in Rome and the desire not to antagonize those they want to convert."

This account of persecution is part of a general theme of anti-Christian persecution by both Romans and Jews, one that starts with the Pharisee rejection of Jesus's ministry, the cleansing of the Temple, and continues on with his trial before the High Priest, his crucifixion, and the Pharisees' refusal to accept him as the Jewish messiah. This theme plays a significant part in several Christian doctrines ranging from the release of Christians from obeying the Old Testament Law to the commandment to preach to "all nations" (meaning to gentiles as well as Jews) to the concepts of supersessionism.

Luke–Acts contains numerous references to and episodes of persecution against Christians, carried out primarily by Jews and Roman authorities. As historiography, the scholarly consensus is that Luke–Acts presents a skewed picture of the hardships faced by the early church. While evidence for isolated incidents has been found, there exists no historical evidence for systematic persecution of early Christians by either Jews or the Romans. during the period Luke–Acts was likely composed. The alleged historical unreliability of Luke-Acts has not, however, prevented scholars from asking what purpose persecution serves in the narrative and what Luke–Acts' presentation of persecution suggests about the author's motivations. Possible explanations for the purpose of persecution in Luke–Acts have included: to portray Christianity as a non-threat to the Romans by contrasting the movement with a disruptive Jewish community; to craft a polemic to discredit critics of Christianity; and to provide encouragement in times of hardship.

The Gospel of Luke contains a few explicit examples of persecution against Christians. There are several passages that mention coming difficulties for Jesus' followers and Jerusalem, which scholars believe foreshadow the hardships Christians will face in Acts. For instance, while Matthew (13:53-58) and Mark (6:1-6) have versions of the rejection of Jesus in his hometown, Luke (4:14-30) devotes much more time to the episode than the other gospels. S. G. Wilson suggests that this might give a glimpse of later persecution by Jews and rejection of the Jewish mission for a gentile mission in Acts (13:46). In Luke, Jesus speaks of "people [who] hate" and "defame you on account of the Son of Man" and likens his followers' suffering to that of earlier prophets (6:22-23 NRSV). Jesus later says to "not fear those who kill the body and after that can do nothing more" (12:4 NRSV).

The major exception to these less direct references to persecution is the Passion narrative in 22:1-23:56. Jewish chief priests and scribes plot to kill Jesus (22:1-6), arrest him (22:47-52), question him before the Sanhedrin and then take him to the Roman prefect Pontius Pilate (22:66-23:1). Jesus is questioned by Pilate (23:3-5), "Herod" (believed to be Agrippa I) (23:6-12), sentenced to death (23:21-25), crucified, died, and buried (23:26-56). Luke's depiction of Pilate is thought by scholars to be key to understanding the author of Luke–Acts' opinion of the Roman Empire.

The Acts of the Apostles, which tells the story of the early Christian church, contains a multitude of episodes highlighting persecution. The majority of these conflicts occur between Christians and Jews, though there are examples of persecution by gentiles (such as the diviner-slave's owners in 16:16-24) and Romans (28:16). While Kelhoffer and Wilson have argued there is a purposeful pattern of Jewish-led persecution in Luke–Acts, there is significant doubt over how historically accurate Luke–Acts' portrayal of Christian-Jewish relations is and how wide reaching (whether directed at specific Jewish groups or 'Jews' as a whole) and seriously this polemic was meant be taken.

The Roman court system features heavily in the later chapters when Saint Paul is brought to trial before several different officials. Paul is tried through the procedure of cognitio extra ordinem, wherein the Roman magistrate participates in all parts of the trial, from evidence gathering to inquisition to judgment. A similar system can be seen in Pliny the Younger's letter 10.96