Shrine of Our Lady of Guadalupe is a Catholic shrine located in La Crosse, Wisconsin. It is dedicated to the Blessed Virgin Mary under the title of Our Lady of Guadalupe. The Construction of the Shrine Church began on May 13, 2004, with a dedication on July 31, 2008. The 100-acre (0.40 km2) grounds include a visitors' center and outdoor devotional areas such as a rosary walk, Stations of the Cross, and a votive candle chapel. The shrine was founded and later dedicated by Archbishop Raymond Leo Burke. Mass and the sacrament of Penance are celebrated daily by Franciscan Friars of the Immaculate.

The shrine was an inspiration of Cardinal Raymond Leo Burke, while he was Bishop of La Crosse, who wanted to establish a place of lasting worship for Roman Catholics to go on pilgrimage to in the Diocese of La Crosse. On September 28, 1999 a letter was sent for consideration to the Vatican, and on November 11, 1999 the Holy See gave the project its approval and blessing. 70 acres (280,000 m2) of woodland near the south end of La Crosse were then donated by Robert and Lucille Swing. Groundbreaking began on June 17, 2001.

The first phase of construction included a Pilgrim Center, which features an orientation room, information desk, the Flores Mariae gift shop, and the Culina Mariana restaurant, and the Mother of Good Counsel Votive Candle Chapel. This phase was completed and dedicated December 12, 2002.

The second phase included an outdoor stations of the cross, a devotional area to St. Joseph the Workman and a rosary walk, in addition to construction on the Shrine Church. Groundbreaking for the church was on May 13, 2004. The Stations of the Cross were dedicated December 9, 2004, followed by the devotional area to St. Joseph the Workman on September 21, 2007 and the rosary walk on December 8, 2007.

Excavation for the third phase, the Memorial to the Unborn, began on October 29, 2007.

On July 31, 2008 the Shrine Church was dedicated. The dedication Mass was also presided over by Burke, who by then had been named Prefect of the Supreme Tribunal of the Apostolic Signatura, having previously served as Archbishop of Saint Louis and before that as Bishop of La Crosse, and who would be named a Cardinal in 2010. Burke was joined by Cardinals Justin Francis Rigali of Philadelphia, who had previously served as Archbishop of Saint Louis before Burke was named as his successor, and Francis Eugene George of Chicago, the closest cardinalatial see to La Crosse. The ceremony included an honor guard made up of members of the local area Knights of Columbus and Knights and Ladies of the Holy Sepulchre. Also in attendance were over 100 priests, and members of the St. Juan Diego Guild and Marian Catechist Apostolate.

The Memorial to the Unborn was completed on December 12, 2008 and was also dedicated by Archbishop Burke.