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Article
Enzymatic construction of shRNA library from oligonucleotide library
Short hairpin RNAs (shRNAs) expressed from vectors have been used as an effective means of exploiting the RNA interference (RNAi) pathway in mammalian cells. Genome-scale screening with shRNA libraries has bee...
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Article
Promoter cross-talk affects the inducible expression of intronic shRNAs from the tetracycline response element
RNA interference (RNAi), defined as double-stranded, RNA-mediated gene silencing, is a useful tool for functional genomic studies. Along with increasing information about genomic sequences due to the innovativ...
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Article
Enhancement of gene knockdown efficiency by CNNC motifs in the intronic shRNA precursor
Short hairpin RNAs (shRNAs) expressed from vectors have been used as an effective means of exploiting the RNA interference (RNAi) pathway in mammalian cells. Of several methods to express shRNA, a method of tr...
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Article
Open AccessNanobody-targeted E3-ubiquitin ligase complex degrades nuclear proteins
Targeted protein degradation is a powerful tool in determining the function of specific proteins or protein complexes. We fused nanobodies to SPOP, an adaptor protein of the Cullin-RING E3 ubiquitin ligase com...
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Article
Enzymatic production and expression of shRNAmir30 from cDNAs
RNA interference (RNAi) is an important tool for studying gene function and genetic networks. Double-stranded RNA (dsRNA) triggers RNAi that selectively silences gene expression mainly by degrading target mRNA...
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Article
Live image profiling of neural crest lineages in zebrafish transgenic lines
Zebrafish transgenic lines are important experimental tools for lineage tracing and imaging studies. It is crucial to precisely characterize the cell lineages labeled in transgenic lines to understand their li...
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Article
LIN-12/notch regulates lag-1 and lin-12 expression during anchor cell/ventral uterine precursor cell fate specification
During Caenorhabditis elegans gonadal development, a stochastic interaction between the LIN-12/Notch receptor and the LAG-2/Delta ligand initiates cell fate specification of two equivalent pre-anchor cell (AC)/pr...
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Article
Regulatory regions of the C elegans genome contain more low-affinity REF-1 transcription-factor binding sites than high-affinity sites
Regulatory regions that control transcription generally contain multiple binding sites for sequence-specific, DNA-binding proteins. Such regions are characterized by clusters of cis-acting sites that are more evo...
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Protocol
Electrophoretic Mobility Shift Assays for Protein–DNA Complexes Involved in DNA Repair
The electrophoretic mobility shift assay (EMSA) can be used to study proteins that bind to DNA structures created by DNA-damaging agents. UV-damaged DNA-binding protein (UV-DDB), which is involved in nucleotid...
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Protocol
Electrophoretic Mobility Shift Assays to Study Protein Binding to Damaged DNA
The electrophoretic mobility shift assay (EMSA) can be used to identify proteins that bind specifically to damaged DNA. EMSAs detect the presence of key DNA repair proteins, such as ultraviolet (UV)-damaged DN...
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Protocol
The Use of Electrophoretic Mobility Shift Assays to Study DNA Repair
DNA repair pathways must include proteins that recognize and bind to damaged DNA. The search for such proteins has been facilitated by the use of electrophoretic mobility shift assays (EMSAs), which were first...
