Category Ranking
98%
Total Visits
921
Avg Visit Duration
2 minutes
Citations
20
Article Abstract
Fluorescence in situ hybridization (FISH) has been widely used to visualize target DNA sequences in fixed chromosome samples by denaturing the dsDNA to allow complementary probe hybridization, thus damaging the chromatin structure by harsh treatments. To overcome this limitation, a CRISPR/Cas9-based in situ labeling method was developed, termed CRISPR-FISH. This method is also known as RNA-guided endonuclease-in situ labeling (RGEN-ISL). Here we present different protocols for the application of CRISPR-FISH on acetic acid: ethanol or formaldehyde-fixed nuclei and chromosomes as well as tissue sections for labeling repetitive sequences in a range of plant species. In addition, methods on how immunostaining can be combined with CRISPR-FISH are provided.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-0716-3226-0_20 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Transition of Structurally Distinct Amyloids in the Degradation of Protein Materials.
J Phys Chem B
September 2025
Chemistry Division, Code 6176, US Naval Research Laboratory, Washington, D.C. 20375, United States.
Amyloid materials are formed from the aggregation of single proteins, yet contain polymorphisms where bulk properties are defined by a composition of multiple fibril types. Though desirable as a sustainable material, little is known about how various fibril types survive at high temperatures or in nonpolar solvents due to their highly similar molecular and nanoscale features. Here, we demonstrate that in situ two-dimensional infrared spectroscopy (2DIR), when paired with nanoscale microscopy, can determine the transition temperature of amyloid subpopulations without the use of labels.
View Article and Find Full Text PDFSingle-Cell Membrane Molecular Cartography Enabled by Nanoengineered VUV-LDI Mass Spectrometry Imaging.
Anal Chem
September 2025
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China.
Deciphering the multicomponent of cell membranes at the single-cell level is critical for understanding pathological mechanisms such as tumor metastasis, yet remains technically daunting due to the membrane's nanoscale thickness and ultralow molecular abundance. Here, we introduce a surface-assisted vacuum ultraviolet laser desorption-ionization mass spectrometry imaging (SAVUVDI-MSI) platform that overcomes long-standing challenges of cytoplasmic interference and insufficient sensitivity. Leveraging the nanoscale depth profiling capability of VUV-LDI, we achieve precise ablation of a single-cell membrane.
View Article and Find Full Text PDFVisible Light-Driven Benzylic C(sp)-H Carboxylation Enables Synthesis of C-Labeled (±)-α-Amino Acids with C-Formate.
Org Lett
September 2025
Frontiers Science Center for Transformative Molecules, State Key Laboratory of Polyolefins and Catalysis, State Key Laboratory of Synergistic Chem-Bio Synthesis, Zhang Jiang Institute for Advanced Study, Shanghai Jiao Tong University, Shanghai 200240, China.
C-labeled α-amino acids are important molecules in biological studies and drug development. Cost-effective synthesis of α-amino acids with a high level of C incorporation under mild conditions remains limited. Herein, we report the development of a benzylic C(sp)-H carboxylation method to prepare highly C-labeled α-amino acids, i.
View Article and Find Full Text PDFImaging of Anodic Electrocatalytic Reactions via Cathodic Electrochemiluminescence on Focused Ion Beam-Fabricated Closed Bipolar Nanoelectrode Arrays.
Anal Chem
September 2025
Department of Chemistry, Southern University of Science and Technology, 518055 Shenzhen, China.
Electrochemiluminescence (ECL) imaging through closed bipolar nanoelectrode arrays (BPnEAs) has emerged as a promising method for in situ label-free wide-field electrochemical imaging. In this study, a cathodic ECL system based on [Ru(bpz)]/SO is combined with the BPnEAs fabricated on silicon nitride membrane windows through focused ion beam nanofabrication, enabling effective bipolar imaging of heterogeneous anodic electrocatalytic reactions. The shape, distribution, size, and material composition of individual electrodes within the array can be precisely controlled.
View Article and Find Full Text PDFGenetic Code Expansion-Driven in Situ Click Labeling Enables Rapid Imaging-Based Selection of Functional Nanobody-Dye Conjugates.
Chem Pharm Bull (Tokyo)
September 2025
Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Antigen-binding proteins, such as nanobodies, modified with functional small molecules hold great potential for applications including imaging probes, drug conjugates, and localized catalysts. However, traditional chemical labeling methods that randomly target lysine or cysteine residues often produce heterogeneous conjugates with limited reproducibility. Conventional site-specific conjugation approaches, which typically modify only the N- or C-terminus, may also be insufficient to achieve the desired functionalities.
View Article and Find Full Text PDF