These cell lines are part of an effort funded by NIH R24 OD019847 (N. Perrimon, PI) to develop a research resource. Specifically, the Drosophila RNAi Screening Center (DRSC) at Harvard Medical School developed a set of GFP-tagged cell lines using CRISPR knock-in technology, then shared the cell lines with the Drosophila Genome Resource Center (DGRC) for distribution to the community. The parental cell line is the DRSC cell line S2R+-MT::Cas9 (DGRC stock number 268). This parental cell line is itself a derivative of NPT005, which is positive for attP and mCherry-Clic, modified by stable transfection of Cas9. We further modified DRSC S2R+-MT::Cas9 by using a donor and sgRNA to knock GFP into an endogenous locus. All GFP-modified cell lines donated by the DRSC to the DGRC were single-cell cloned and verified by imaging and molecular analysis prior to deposition into the DGRC collection. For detailed information about the endogenous target sites and corresponding validation data for each cell line in the collection, please refer to the associate file spreadsheet and its embedded files. Most of these cell lines were made by the DRSC (S. Knight, D. Yang-Zhou, J. Zirin, S.E. Mohr, N. Perrimon) in collaboration with O. Kanca in the Bellen lab (Baylor College of Medicine). It is important to note that not all alleles have been modified, such that the GFP tag could be lost after repeated passaging. We recommend using fluorescence-activated cell sorting (FACS) to assess and recover GFP-positive cells. If use of cell lines from this collection results in publication, please acknowledge NIH R24 OD019847.