That lead is held by one of her rivals for the world number one ranking, Thailand's Ariya Jutanugarn.
Jutanugarn had a bogey-free six-under 66 to move to nine-under for the tournament, sharing the lead with the Korean duo of In-Kyung Kim and Su-Yeon Jang.
Jutanugarn won five times last season and was the LPGA Tour's player of the year, and can continue to cut into Ko's lead on top of the world rankings if she wins this weekend.
Likewise world number two So Yeon Ryu is also a shot at snaring the world number one ranking, and sits just two shots off the lead in a share of eighth place.
Ko has been world number one for 76 weeks, but has had a large lead slowly chipped away by the chasing duo. However, her lead is still sizeable enough that both Ryu and Jutanugarn would need to win, and have Ko finish poorly, to end her reign atop the rankings.
Fellow Kiwi golfer Danny Lee has started strongly on the PGA Tour at the RBC Heritage event. Lee shot a five-under par to sit in a share of fifth, three shots off the lead held by American Bud Cauley.
"My putting was falling nicely all day," said Lee. "It's a very tricky course, but it was perfect conditions for scoring."
Lee has recorded three top 25 finishes in his last three four starts, but is yet to record a top 15 finish in 2017.