cellprofiler worm toolbox python source code

CELLPROFILER WORM TOOLBOX PYTHON SOURCE CODE DRIVER
CELLPROFILER WORM TOOLBOX PYTHON SOURCE CODE FULL

CELLPROFILER WORM TOOLBOX PYTHON SOURCE CODE DRIVER

RNAi can be induced (B) by bathing cells in aqueous dsRNA solution (C) by microinjections of dsRNA into embryos, (D) by crossing of transgenic (Gal4) driver lines to dsRNA-expressing flies (UAS-dsRNA), or (E) shRNA-expressing flies (UAS-shRNA). RISC comprises the siRNA, AGO2 (green), and other accessory proteins ( e.g., hsp90, blue) and binds and degrades the siRNA complementary target mRNA (red). Each dsRNA/shRNA molecule is then processed by Dicer-2 and R2D2 (brown) into multiple ∼19-bp single-stranded siRNAs. (A) In Drosophila cells, dsRNAs (black) are taken up by cells using “scavenger” receptor-mediated endocytosis. RNAi is a gene silencing method that works through degradation of homologous messenger RNAs (mRNA, orange). 2006 Pratt and MacRae 2009 Iwasaki et al. The loaded RISC can also interact with nonintended homologous target sequences, such as near-perfect matches in 3′-UTRs, leading to miRNA-like inhibition of translation, which can be a major source of off-target effects ( Hannon 2002 Kulkarni et al.

Endogenous small RNAs such as micro RNAs (miRNAs) use similar and divergent pathways to silence gene expression. This complex identifies sequence-homologous endogenous RNAs through a homology-seeking activity, leading to their cleavage and degradation.

CELLPROFILER WORM TOOLBOX PYTHON SOURCE CODE FULL

Subsequently, the full RNA-induced silencer complex (RISC) is formed. This in turn induces the association of the argonaute protein Ago2, which is stabilized by a HSC70/Hsp90 chaperone system, and then leads to the unwinding of the duplex, its cleavage, and finally ejection of the passenger strand ( Iwasaki et al. Mechanistic studies, mainly performed in Drosophila, elucidated a biochemical pathway that upon introduction of exogenous dsRNA leads to the formation of a complex, consisting of Dicer-2 and R2D2, which cuts the duplex RNAs into short 21-nucleotide (nt)-long fragments ( Figure 1A). RNAi was first discovered in Caenorhabditis elegans, where it was shown that injection of long dsRNA or feeding worms with bacteria expressing dsRNA is sufficient to silence gene expression, leading to highly penetrant phenotypes ( Fire et al. RNA interference (RNAi) is an endogenous cellular mechanism triggered by double-stranded RNA (dsRNA), which leads to the degradation of homologous RNAs. Furthermore, we will review the generation and use of genome-scale RNAi libraries for tissue-specific knockdown analysis in vivo and discuss the differences and similarities with the use of genome-engineering methods such as CRISPR/Cas9 for functional analysis. We will discuss available reagent libraries and cell lines, methodological approaches for cell-based assays, and computational methods for the analysis of high-throughput screens. In this review, we will describe the use of RNAi to study gene function in Drosophila with a particular focus on high-throughput screening methods applied in cultured cells. In Drosophila, RNAi has been applied in cultured cells or in vivo to perturb the function of single genes or to systematically probe gene function on a genome-wide scale. First discovered in Caenorhabditis elegans, RNAi can be used to silence the expression of genes through introduction of exogenous double-stranded RNA into cells.

In the last decade, RNA interference (RNAi), a cellular mechanism that uses RNA-guided degradation of messenger RNA transcripts, has had an important impact on identifying and characterizing gene function.

Cellprofiler worm toolbox python source code