It happened around 2:30 a.m. near the Academy Road exit at mile marker 32.
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Police say three men, who were all in the same vehicle, were injured in the shooting that shut down the highway for hours.
According to state police, a 25-year-old man from Trenton was shot and taken to the hospital in critical condition.
Two other victims -- a 25-year-old man from Morrisville, Pa. and a 29-year-old man from Trenton -- are both listed as stable. The Morrisville victim suffered a graze wound, police said.
All northbound lanes of I-95 between Exit 26/Betsy Ross Bridge to New Jersey and Exit 32/Academy Road/Linden Ave were closed for several hours as many hit the road for the holiday.
Traffic was being forced off the northbound side of the interstate at Aramingo Avenue, causing major delays.
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"As a rider, it's definitely something to be concerned about. It may be hard to be solved, but it's definitely a very concerning issue in the city," said Rachshawn Daniels of Frankford.
This is the latest shooting on a highway in the Philadelphia area. In April, part of the I-76 was shut down after a man was shot in the neck.
On May 1, on I-95 north, police say a driver fired at another car, grazing the victim. In June, an apparent case of road rage led to a shooting on I-95 near Prospect Park.
Just a couple of weeks ago, a shooting investigation closed off a stretch of I-95 in Delaware County.
State police say they are looking for another car that could have been involved.
Further details, including what led up to the gunfire, were not immediately released.