title: An Efficient Method for Electroporation of Small Interfering RNAs into ENCODE Project Tier 1 GM12878 and K562 Cell Lines.
abstract: The Encyclopedia of DNA Elements (ENCODE) Project aims to identify all functional sequence elements in the human genome sequence by use of high-throughput DNA/cDNA sequencing approaches. To aid the standardization, comparison, and integration of data sets produced from different technologies and platforms, the ENCODE Consortium selected several standard human cell lines to be used by the ENCODE Projects. The Tier 1 ENCODE cell lines include GM12878, K562, and H1 human embryonic stem cell lines. GM12878 is a lymphoblastoid cell line, transformed with the Epstein-Barr virus, that was selected by the International HapMap Project for whole genome and transcriptome sequencing by use of the Illumina platform. K562 is an immortalized myelogenous leukemia cell line. The GM12878 cell line is attractive for the ENCODE Projects, as it offers potential synergy with the International HapMap Project. Despite the vast amount of sequencing data available on the GM12878 cell line through the ENCODE Project, including transcriptome, chromatin immunoprecipitation-sequencing for histone marks, and transcription factors, no small interfering siRNA-mediated knockdown studies have been performed in the GM12878 cell line, as cationic lipid-mediated transfection methods are inefficient for lymphoid cell lines. Here, we present an efficient and reproducible method for transfection of a variety of siRNAs into the GM12878 and K562 cell lines, which subsequently results in targeted protein depletion.
authors.affiliation: 1 Center for RNA Systems Biology, 2 California Institute for Quantitative Biosciences (QB3), and Departments of 3 Chemistry and 4 Molecular and Cell Biology, University of California, Berkeley, California, USA.
authors.forename: Ryan Y
authors.initials: RY
authors.lastname: Muller
authors.affiliation: 1 Center for RNA Systems Biology, 2 California Institute for Quantitative Biosciences (QB3), and Departments of 3 Chemistry and 4 Molecular and Cell Biology, University of California, Berkeley, California, USA.
authors.forename: Ming C
authors.initials: MC
authors.lastname: Hammond
authors.affiliation: 1 Center for RNA Systems Biology, 2 California Institute for Quantitative Biosciences (QB3), and Departments of 3 Chemistry and 4 Molecular and Cell Biology, University of California, Berkeley, California, USA.
authors.forename: Donald C
authors.initials: DC
authors.lastname: Rio
authors.affiliation: 1 Center for RNA Systems Biology, 2 California Institute for Quantitative Biosciences (QB3), and Departments of 3 Chemistry and 4 Molecular and Cell Biology, University of California, Berkeley, California, USA.
authors.forename: Yeon J
authors.initials: YJ
authors.lastname: Lee
mesh_terms: D053487:DEAD-box RNA Helicases / Q000096:biosynthesis / Q000235:genetics; D018274:Electroporation; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D020014:K562 Cells; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection / Q000379:methods*
chemical_list: D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D034741:RNA, Small Interfering; C506749:Ddx5 protein, human; D053487:DEAD-box RNA Helicases
keywords: RNAi studies; protein knockdown; siRNA; transfection