Saturday, September 27, 2014

Drosophila Models of Human Disease: Large-scale study of genes on the fly X-chromosome...

Drosophila Models of Human Disease: Large-scale study of genes on the fly X-chromosome...: Yamamoto S, Jaiswal M, Charng WL, Gambin T, Karaca E, Mirzaa G, Wiszniewski W, Sandoval H, Haelterman NA, Xiong B, Zhang K, Bayat V, David G...

Study related to Abl kinase uses primary cultured cells

This is not an RNAi study but the approach taken might of interest to those thinking about cell-based studies. Liu L, Wu CF. Distinct effects of Abelson kinase mutations on myocytes and neurons in dissociated Drosophila embryonic cultures: mimicking of high temperature. PLoS One. 2014 Jan 21;9(1):e86438. PMID: 24466097; PMCID: PMC3897706. From the abstract: "... we observed myocyte fusion events and nerve-muscle contact formation between WT and Abl cells in mixed WT and Abl cultures derived from labeled embryos ..."

Protocol paper -- RNAi and other techniques in gut-microbe studies

Houtz PL, Buchon N. Methods to Assess Intestinal Stem Cell Activity in Response to Microbes in Drosophila melanogaster. Methods Mol Biol. 2014;1213:171-82. PMID: 25173382.

Protocol paper includes information about RNAi in the germline

DeVorkin L, Gorski SM. Genetic manipulation of autophagy in the Drosophila ovary. Cold Spring Harb Protoc. 2014 Sep 2;2014(9):pdb.prot080358. PMID: 25183818. From the abstract: "... Here we describe how to generate a homozygous mutant germline using the FLP-DFS (dominant female sterile) technique, how to generate somatic clones, and how to induce targeted gene knockdown in the germline using RNAi."

Sunday, September 7, 2014

UAS system combined with CRISPR-Cas9 in Drosophila

Zhaoyu Xue, Menghua Wu, Kejia Wen, Menda Ren, Li Long, Xuedi Zhang and Guanjun Gao. CRISPR/Cas9 Mediates Efficient Conditional Mutagenesis in Drosophila. G3. Available as early online content.

From the abstract:  "... tissue-specific expression of Cas9 driven by the Gal4/upstream activating site (UAS) system with various ubiquitously expressed gRNA transgenes to effectively inactivate gene expression in a temporally and spatially controlled manner. ... by including multiple gRNAs in a transgenic vector to target a single gene, we achieved a high degree of gene mutagenesis in specific tissues. ..."

Thursday, September 4, 2014

Genome editing--design tools

On my desk to read today--another paper related to CRISPRs.

Montague TG, Cruz JM, Gagnon JA, Church GM, Valen E. CHOPCHOP: a CRISPR/Cas9 and TALEN web tool for genome editing. Nucleic Acids Res. 2014 Jul;42(Web Server issue):W401-7. PMID: 24861617.

This report describes the ChopChop suite of tools, which includes support for Drosophila sgRNA design.

The DRSC offers our CRISPR2 sgRNA design tool, which provides a genome-browse view and includes efficiency scores (or you can run any designs through a 'calculator').

Additional tools are listed at AddGene.

The DRSC offers support for sgRNA and donor construct building through the Genome Engineering Production Group.

Wednesday, September 3, 2014

RNAi screening in fly cells to investigate Brucella infection

Pandey A, Ding SL, Ficht TA, de Figueiredo P. siRNA Screens Using Drosophila Cells to Identify Host Factors Required for Infection. Methods Mol Biol. 2014;1197:229-44. PMID: 25172284.