The Carnegie Protein Trap Library: A Versatile Tool for Drosophila Developmental Studies

Abstract

Metazoan physiology depends on intricate patterns of gene expression that remain poorly known. Using transposon mutagenesis in Drosophila, we constructed a library of 7404 protein trap and enhancer trap lines, the Carnegie collection, to facilitate gene expression mapping at single-cell resolution. By sequencing the genomic insertion sites, determining splicing patterns downstream of the enhanced green fluorescent protein (EGFP) exon, and analyzing expression patterns in the ovary and salivary gland, we found that 600-900 different genes are trapped in our collection. A core set of 244 lines trapped different identifiable protein isoforms, while insertions likely to act as GFP-enhancer traps were found in 256 additional genes. At least 8 novel genes were also identified. Our results demonstrate that the Carnegie collection will be useful as a discovery tool in diverse areas of cell and developmental biology and suggest new strategies for greatly increasing the coverage of the Drosophila proteome with protein trap insertions.

Loss of lysine-specific demethylase 1 nonautonomously causes stem cell tumors in the Drosophila ovary

“…Cap cells and terminal filament cells express high levels of Lamin C and hh (7,27). CB03410, a previously identified protein trap line (33), is also expressed in adult cap cells and terminal filament cells. In Lsd1 mutants, the expression of all three markers expanded to most of the somatic cells within the germarium (Fig.…”

Opening Pathways for Underrepresented High School Students to Biomedical Research Careers: The Emory University RISE Program

“…The research project carried out by students in the RISE program seeks to find novel protein components of insulator complexes as well as other proteins that may have unique roles in nuclear organization. The approach is to screen a collection of GFP-tagged protein-trap Drosophila alleles to identify those with interesting nuclear localization patterns (Morin et al 2001;Kelso et al 2004;Buszczak et al 2007). Once a protein is identified, further analysis is carried out to elucidate its role in chromatin organization and nuclear biology.…”

Requirements of Lgl in cell differentiation and motility during Drosophila ovarian follicular epithelium morphogenesis

“…[5][6][7][8][9][10][11][12][13][14] In addition, the apico-basal polarity and DE-Cadherin based adhesion are crucial for the integrity of border cell cluster during its morphogenetic movement. 6,15,16 As developmental cell migration and pathological cell invasion involve common cellular and biological processes, 3,10,[17][18][19][20][21] further investigation of this model system will contribute to unraveling the molecular mechanisms underlying cancer invasion and metastasis. The Drosophila nTSGs lgl, dlg and scrib cooperatively function in multiple biological processes, including cell polarity and proliferation control in epithelial tissues, tumor cell invasion and asymmetric division of neuroblasts.…”