As PLOS Biology turns 20 years old, we are celebrating two decades of publishing high-impact life sciences research at the forefront of open science. This collection contains articles that look back at landmark PLOS Biology studies and how they have influenced their respective fields, and others that look at how far given fields have come in the past 20 years and where they are going, and others that discuss how publishing and open science have evolved in the past 20 years and what is to come. We will continue to update this collection with content throughout 2023.