Among those in her delegation are Rep. Keith Ellison, the first Muslim member of Congress.
Pelosi's repeat trip to the Middle East indicates she has no intention of letting the White House have the sole province on foreign policy. She has already forced legislation through the House that would order all combat troops out of Iraq by September 2008, a measure that resembles a similar measure approved by the Democratic-run Senate.
The Bush administration has mostly refused to engage Syria diplomatically because of its ties to terrorist networks. U.S. officials held their first direct, high-level contact with Syrian representatives in years this month when they met with officials from several Middle East countries in Baghdad to discuss Iraq.
Others traveling with Pelosi include Rep. Tom Lantos, the Democratic chairman of the House Foreign Affairs Committee
The House has adjourned for a two-week spring break.
The group planned to meet with Israeli Prime Minister Ehud Olmert, and to travel to the West Bank to meet with Palestinian President Mahmoud Abbas, said Ellison's spokesman, Rick Jauert.
The speaker plans to address the Israeli Knesset on Sunday in what will be her first address to a foreign government legislature and as the highest ranking American woman to speak before the Israeli parliament, according to Pelosi's office.
She is expected to discuss "America's commitment to Israel and the challenges facing the two nations in the Middle East," according to a statement.
In late January, Pelosi led a delegation of House members to Iraq, Afghanistan, Israel and other neighboring countries.
The January trip to Baghdad came just days after the president asked Congress in his State of the Union address to give his revised war strategy a chance to work. The United States is sending more than 21,500 additional combat troops, plus thousands of other support troops, to Iraq in a bid to tamp down sectarian attacks and provide enough security to hasten reconstruction efforts.