Genome editing comes of age.

abstract：:High throughput microarray transcription analyses provide us with the expression profiles for large amounts of plant genes. However, their tissue and cellular resolution is limited. Thus, for detailed functional analysis, it is still necessary to examine the expression pattern of selected candidate genes at a cellular...

journal_title：Nature protocols

authors： Hejátko J,Blilou I,Brewer PB,Friml J,Scheres B,Benková E

更新日期：2006-01-01 00:00:00

abstract：:Fluorescent microspheres are commonly used to assess bone blood supply in large animals, but the technique is not widely used in smaller mammals, as traditional methods such as reference blood sampling, ventilation and catheterization are not easily applied. This protocol describes a viable alternative for measuring b...

journal_title：Nature protocols

authors： Serrat MA

更新日期：2009-01-01 00:00:00

abstract：:A procedure for the synthesis of N-succinimidyl 3-iodobenzoate labeled with any iodine isotope ([*I]SIB), which is an agent used in the radioiodination of proteins and peptides, from its tin precursor N-succinimidyl 3-(tri-n-butylstannyl)benzoate (STB) is described. Also included are protocols for the synthesis of an ...

journal_title：Nature protocols

authors： Vaidyanathan G,Zalutsky MR

更新日期：2006-01-01 00:00:00

abstract：:Optogenetic tools provide users the ability to photocontrol the activity of cells. Commonly, activation is achieved by expression of proteins from photosynthetic organisms, for example, microbial opsins (e.g., ChR2). Alternatively, a sister approach, synthetic optogenetics, enables photocontrol over proteins of mammal...

journal_title：Nature protocols

authors： Carmi I,De Battista M,Maddalena L,Carroll EC,Kienzler MA,Berlin S

更新日期：2019-03-01 00:00:00

abstract：:A big challenge in proteomics is the identification of cell-type-specific proteomes in vivo. This protocol describes how to label, purify and identify cell-type-specific proteomes in living mice. To make this possible, we created a Cre-recombinase-inducible mouse line expressing a mutant methionyl-tRNA synthetase (L27...

journal_title：Nature protocols

authors： Alvarez-Castelao B,Schanzenbächer CT,Langer JD,Schuman EM

更新日期：2019-02-01 00:00:00

abstract：:Isolated protoplasts serve as a transient expression system that is highly representative of stable transgenics in terms of transcriptome responses. They can also be used as a cellular system to study gene transactivation and nucleocytoplasmic protein trafficking. They are particularly useful for systems studies in wh...

journal_title：Nature protocols

authors： Lin YC,Li W,Chen H,Li Q,Sun YH,Shi R,Lin CY,Wang JP,Chen HC,Chuang L,Qu GZ,Sederoff RR,Chiang VL

更新日期：2014-09-01 00:00:00

abstract：:Growing evidence indicates that RNA G-quadruplexes have important roles in various processes such as transcription, translation, regulation of telomere length, and formation of telomeric heterochromatin. Investigation of RNA G-quadruplex structures associated with biological events is therefore essential to understand...

journal_title：Nature protocols

authors： Bao HL,Xu Y

更新日期：2018-04-01 00:00:00

abstract：:The goal of many current studies of neovascularization is to define the phenotype of vascular cell populations of different origins and to determine how such cells promote assembly of vascular channel. Here, we describe a protocol to immunophenotype vascular cells by high-resolution imaging and by fluorescence-activat...

journal_title：Nature protocols

authors： Jones RC,Capen DE,Cohen KS,Munn LL,Jain RK,Duda DG

更新日期：2008-01-01 00:00:00

abstract：:Raman spectroscopy can be used to measure the chemical composition of a sample, which can in turn be used to extract biological information. Many materials have characteristic Raman spectra, which means that Raman spectroscopy has proven to be an effective analytical approach in geology, semiconductor, materials and p...

journal_title：Nature protocols

authors： Butler HJ,Ashton L,Bird B,Cinque G,Curtis K,Dorney J,Esmonde-White K,Fullwood NJ,Gardner B,Martin-Hirsch PL,Walsh MJ,McAinsh MR,Stone N,Martin FL

更新日期：2016-04-01 00:00:00

abstract：:In this protocol we describe how to perform the Multi-Source Interference Task (MSIT), a validated functional magnetic resonance imaging (fMRI) task that reliably and robustly activates the cingulo-frontal-parietal cognitive/attention network (CFP network) within individual subjects. The MSIT can be used to (i) identi...

journal_title：Nature protocols

authors： Bush G,Shin LM

更新日期：2006-01-01 00:00:00

abstract：:Here we describe a protocol for the production of frozen competent yeast cells that can be transformed with high efficiency using the lithium acetate/single-stranded carrier DNA/PEG method. This protocol allows the production of highly competent yeast cells that can be frozen and used at a later date and is especially...

journal_title：Nature protocols

authors： Gietz RD,Schiestl RH

更新日期：2007-01-01 00:00:00

abstract：:As proteins within cells are spatially organized according to their role, knowledge about protein localization gives insight into protein function. Here, we describe the LOPIT technique (localization of organelle proteins by isotope tagging) developed for the simultaneous and confident determination of the steady-stat...

journal_title：Nature protocols

authors： Sadowski PG,Dunkley TP,Shadforth IP,Dupree P,Bessant C,Griffin JL,Lilley KS

更新日期：2006-01-01 00:00:00

abstract：:Here, we describe a simple micromolding method to construct three-dimensional arrays of organotypic epithelial tissue structures that approximate in vivo histology. An elastomeric stamp containing an array of posts of defined geometry and spacing is used to mold microscale cavities into the surface of type I collagen ...

journal_title：Nature protocols

authors： Nelson CM,Inman JL,Bissell MJ

更新日期：2008-01-01 00:00:00

abstract：:The plant cell wall is a chemically complex structure composed mostly of polysaccharides. Detailed analyses of these cell wall polysaccharides are essential for our understanding of plant development and for our use of plant biomass (largely wall material) in the food, agriculture, fabric, timber, biofuel and biocompo...

journal_title：Nature protocols

authors： Pettolino FA,Walsh C,Fincher GB,Bacic A

更新日期：2012-09-01 00:00:00

abstract：:Large-insert BAC (bacterial artificial chromosome) and BIBAC (binary BAC) libraries are essential for modern genomics research for all organisms. We helped pioneer the BAC and BIBAC technologies, and by using them we have constructed hundreds of BAC and BIBAC libraries for different species of plants, animals, marine ...

journal_title：Nature protocols

authors： Zhang HB,Scheuring CF,Zhang M,Zhang Y,Wu CC,Dong JJ,Li Y

更新日期：2012-02-16 00:00:00

abstract：:N-Succinimidyl 3-(di-tert-butyl[(18)F]fluorosilyl)benzoate ([(18)F]SiFB) is a highly reactive prosthetic group for radiolabeling of proteins for use in positron emission tomography (PET). It is similar to N-succinimidyl-4-[(18)F]fluorobenzoate ([(18)F]SFB), the 'gold-standard' prosthetic group for protein (18)F-labeli...

journal_title：Nature protocols

authors： Kostikov AP,Chin J,Orchowski K,Schirrmacher E,Niedermoser S,Jurkschat K,Iovkova-Berends L,Wängler C,Wängler B,Schirrmacher R

更新日期：2012-11-01 00:00:00

abstract：:Electrochemically active bacteria (EAB) have the ability to transfer electrons to electron acceptors located outside the cell, and they are widely present in diverse environments. In spite of their important roles in geochemical cycles, environmental remediation and electricity generation, so far, only a limited numbe...

journal_title：Nature protocols

authors： Yuan SJ,Li WW,Cheng YY,He H,Chen JJ,Tong ZH,Lin ZQ,Zhang F,Sheng GP,Yu HQ

更新日期：2014-01-01 00:00:00

abstract：:This protocol describes a culture system in which inner-ear sensory tissue is produced from mouse embryonic stem (ES) cells under chemically defined conditions. This model is amenable to basic and translational investigations into inner ear biology and regeneration. In this protocol, mouse ES cells are aggregated in 9...

journal_title：Nature protocols

authors： Koehler KR,Hashino E

更新日期：2014-01-01 00:00:00

abstract：:Methods development in chromatography is a time-consuming, trial-and-error process that requires laborious experimentation. We describe a high-throughput screening (HTS) protocol for the rapid identification of chromatographic steps for protein purification from cell-free expression broths. Broths containing the prote...

journal_title：Nature protocols

authors： Rege K,Heng M

更新日期：2010-03-01 00:00:00

abstract：:Interactions of proteins with DNA mediate many critical nuclear functions. Chromatin immunoprecipitation (ChIP) is a robust technique for studying protein-DNA interactions. Current ChIP assays, however, either require large cell numbers, which prevent their application to rare cell samples or small-tissue biopsies, or...

journal_title：Nature protocols

authors： Dahl JA,Collas P

更新日期：2008-01-01 00:00:00

abstract：:The cytokinesis-block micronucleus cytome assay is a comprehensive system for measuring DNA damage, cytostasis and cytotoxicity. DNA damage events are scored specifically in once-divided binucleated (BN) cells and include (a) micronuclei (MNi), a biomarker of chromosome breakage and/or whole chromosome loss, (b) nucle...

journal_title：Nature protocols

authors： Fenech M

更新日期：2007-01-01 00:00:00

abstract：:Gram-negative and Gram-positive bacteria release a variety of membrane vesicles through different formation routes. Knowledge of the structure, molecular cargo and function of bacterial extracellular vesicles (BEVs) is primarily obtained from bacteria cultured in laboratory conditions. BEVs in human body fluids have b...

journal_title：Nature protocols

authors： Tulkens J,De Wever O,Hendrix A

更新日期：2020-01-01 00:00:00

abstract：:New technologies have recently enabled saturation mutagenesis and functional analysis of nearly all possible variants of regulatory elements or proteins of interest in single experiments. Here we discuss the past, present, and future of such multiplexed (functional) assays for variant effects (MAVEs). MAVEs provide de...

journal_title：Nature protocols

authors： Gasperini M,Starita L,Shendure J

更新日期：2016-10-01 00:00:00

abstract：:Hepatic stellate cells (HSCs) have been identified as the main fibrogenic cell type in the liver. Hence, efforts to understand hepatic fibrogenesis and to develop treatment strategies have focused on this cell type. HSC isolation, originally developed in rats, has subsequently been adapted to mice, thus allowing the s...

journal_title：Nature protocols

authors： Mederacke I,Dapito DH,Affò S,Uchinami H,Schwabe RF

更新日期：2015-02-01 00:00:00

abstract：:The terminal monosaccharide of cell surface glycoconjugates is typically a sialic acid (SA), and aberrant sialylation is involved in several diseases. Several methodological approaches in sample preparation and subsequent analysis using mass spectrometry (MS) have enabled the identification of glycosylation sites and ...

journal_title：Nature protocols

authors： Palmisano G,Lendal SE,Engholm-Keller K,Leth-Larsen R,Parker BL,Larsen MR

更新日期：2010-12-01 00:00:00

abstract：:Rapid evaluation of microbial cell-surface carbohydrates is essential to understanding the mechanisms by which bacteria use glycans to establish pathogenic or symbiotic relationships. Microbial glycan analysis is complicated both by the vast diversity of possible carbohydrate structures and by their dynamic nature. Ba...

journal_title：Nature protocols

authors： Hsu KL,Mahal LK

更新日期：2006-01-01 00:00:00

abstract：:We describe the use of a microfabricated cell culture substrate, consisting of a uniform array of closely spaced, vertical, elastomeric microposts, to study the effects of substrate rigidity on cell function. Elastomeric micropost substrates are micromolded from silicon masters comprised of microposts of different hei...

journal_title：Nature protocols

authors： Yang MT,Fu J,Wang YK,Desai RA,Chen CS

更新日期：2011-02-01 00:00:00

abstract：:Reactive oxygen species (ROS) are continuously produced in the cell as a consequence of aerobic metabolism, and are controlled by several antioxidant mechanisms. An accurate measurement of ROS is essential to evaluate the redox status of the cell, or the effects of molecules with the pro-oxidant or antioxidant activit...

journal_title：Nature protocols

authors： Cossarizza A,Ferraresi R,Troiano L,Roat E,Gibellini L,Bertoncelli L,Nasi M,Pinti M

更新日期：2009-01-01 00:00:00

abstract：:The effect of genetic mutation on phenotype is of significant interest in genetics. The type of genetic mutation that causes a single amino acid substitution (AAS) in a protein sequence is called a non-synonymous single nucleotide polymorphism (nsSNP). An nsSNP could potentially affect the function of the protein, sub...

journal_title：Nature protocols

authors： Kumar P,Henikoff S,Ng PC

更新日期：2009-01-01 00:00:00

abstract：:This protocol describes procedures for performing fluorescence resonance energy transfer (FRET) microscopy analysis by three different methods: acceptor photobleaching, sensitized emission and spectral imaging. We also discuss anisotropy and fluorescence lifetime imaging microscopy-based FRET techniques. By using the ...

journal_title：Nature protocols

authors： Broussard JA,Rappaz B,Webb DJ,Brown CM

更新日期：2013-02-01 00:00:00