The bill failed on a 50-50 roll call vote, needing 60 votes to break a filibuster. Three members of the Democratic caucus and all 47 Republicans voted against the bill.
The Obama administration has stated the bill would support 400,000 teacher jobs, preventing layoffs and allowing governments to rehire educators and first responders who have been let go. The $35 billion bill was the first part of the administration's attempt to break out pieces of his $447 billion jobs proposal into separate elements.
After the vote, Obama released a statement voicing his frustration with Republican obstruction of the proposal.
"For the second time in two weeks, every single Republican in the United States Senate has chosen to obstruct a bill that would create jobs and get our economy going again," the president said in the statement. "That's unacceptable."
Sen. Marc Rubio, R-Fla., summarized the GOP opposition on the bill in a statement of his own.
"I support a real jobs plan. I don't think what we voted on today is a real jobs plan. I think it's another bailout of local governments," Rubio said in the statement. "We cannot afford to be bailing out local governments, and we can't afford stimulus 2.0."