title: Using CRISPR-Cas9 to Study ERK Signaling in Drosophila.
abstract: Genome engineering using the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated nuclease 9 (Cas9) technology is revolutionizing biomedical research. CRISPR-Cas9 enables precise editing of genes in a wide variety of cells and organisms, thereby accelerating molecular studies via targeted mutagenesis, epitope tagging, and other custom genetic modifications. Here, we illustrate the CRISPR-Cas9 methodology by focusing on Capicua (Cic), a nuclear transcriptional repressor directly phosphorylated and inactivated by ERK/MAPK. Specifically, we use CRISPR-Cas9 for targeting an ERK docking site of Drosophila Cic, thus generating ERK-insensitive mutants of this important signaling sensor.
authors.affiliation: Institut de Biologia Molecular de Barcelon-CSIC, Parc Cientific de Barcelona, Baldiri Reixac 10, 08028, Barcelona, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Forés
authors.affiliation: Institut de Biologia Molecular de Barcelon-CSIC, Parc Cientific de Barcelona, Baldiri Reixac 10, 08028, Barcelona, Spain.
authors.forename: Aikaterini
authors.initials: A
authors.lastname: Papagianni
authors.affiliation: Institut de Biologia Molecular de Barcelon-CSIC, Parc Cientific de Barcelona, Baldiri Reixac 10, 08028, Barcelona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Rodríguez-Muñoz
authors.affiliation: Institut de Biologia Molecular de Barcelon-CSIC, Parc Cientific de Barcelona, Baldiri Reixac 10, 08028, Barcelona, Spain. gjcbmc@ibmb.csic.es.|ICREA, Pg. Lluís Companys 23, 08010, Barcelona, Spain. gjcbmc@ibmb.csic.es.
authors.forename: Gerardo
authors.initials: G
authors.lastname: Jiménez
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D064113:CRISPR-Cas Systems; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004249:DNA Damage; D004260:DNA Repair; D004330:Drosophila / Q000235:genetics* / Q000378:metabolism*; D020935:MAP Kinase Signaling System; D009154:Mutation; D059372:Nucleotide Motifs; D010641:Phenotype; D011485:Protein Binding; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems; D020794:Receptor Protein-Tyrosine Kinases
keywords: CRISPR; Capicua; Cas9; Docking site; Drosophila; ERK; MAPK; RTK signaling