100 Result(s)
within
Protocol
**an Qi
Life Sciences
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Protocol
Expression, Purification, and Nanodisc Reconstitution of Connexin-43 Hemichannels for Structural Characterization by Cryo-Electron Microscopy
Connexins are polytopic domain membrane proteins that form hexameric hemichannels (HCs) which can assemble into gap junction channels (GJCs) at the interface of two neighboring cells. The HCs may be involved i...
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Single-Molecule Imaging of the Phase Separation-Modulated DNA Compaction to Study Transcriptional Repression
Liquid-liquid phase separation (LLPS) often induces the formation of biomolecule condensates at the cellular level. The importance of this phenomenon has been demonstrated in many important biological function...
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Modeling Plant Transcription Factor Networks Using ConSReg
Plants have developed complex regulatory programs to respond to various environmental stress such as heat, drought, and cold. Systematic understanding of these biological processes depends on robust constructi...
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Identification of Plant Co-regulatory Modules Using CoReg
Regulatory network is often characterized by complex interactions between transcription factors (TFs) and target genes. The synergistic regulations among multiple TFs may co-induce/co-suppress the expressions ...
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High-Throughput γ-H2AX Assay Using Imaging Flow Cytometry
The γ-H2AX assay is a sensitive and reliable method to evaluate radiation-induced DNA double-strand breaks. The conventional γ-H2AX assay detects individual nuclear foci manually, but is labor-intensive and ti...
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Targeted Activation of Arabidopsis Genes by a Potent CRISPR–Act3.0 System
The CRISPR/Cas system has emerged as a versatile platform for sequence-specific genome engineering in plants. Beyond genome editing, CRISPR/Cas systems, based on nuclease-deficient Cas9 (dCas9), have been repu...
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Delivery of CRISPR-Cas12a Ribonucleoprotein Complex for Genome Editing in an Embryogenic Citrus Cell Line
Clustered regularly interspaced short palindromic repeats (CRISPR) technology is a powerful genome editing tool. Recently developed CRISPR-Cas12a system confers several advantages over CRISPR-Cas9, making it i...
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PAM-Less CRISPR-SpRY Genome Editing in Plants
Engineered SpCas9 variant, SpRY, has been demonstrated to facilitate protospacer adjacent motif (PAM) unrestricted targeting of genomic DNA in various biological systems. Here we describe fast, efficient, and ...
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Base Editing in Poplar Through an Agrobacterium-Mediated Transformation Method
CRISPR-Cas9 systems have revolutionized genome editing in plants and facilitated gene knockout and functional genomic studies in woody plants, like poplar. However, in tree species, previous studies have only ...
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Detection and Analysis of Microtubule Nucleator γ-Tubulin Ring Complex
Golgi-derived microtubules constitute an asymmetrical microtubule network that drives polarized transport of vesicles to support cell polarization and directional migration. Golgi-based microtubule nucleation ...
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Assessment of Mitochondrial Dysfunctions After Sirtuin Inhibition
Posttranslational modifications are important for protein functions and cellular signaling pathways. The acetylation of lysine residues is catalyzed by histone acetyltransferases (HATs) and removed by histone ...
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Expression, Purification, and Structure Determination of Human PTCH1–HH-N Complexes
Patched-1 (PTCH1), a tumor suppressor, serves as the receptor of Hedgehog (HH) ligand and negatively regulates the HH signaling pathway. Mutations of PTCH1 are implicated in many human cancers. Structural inve...
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Generation of Monoclonal iPSC Lines with Stable Cas9 Expression and High Cas9 Activity
CRISPR/Cas9-mediated gene editing has been rapidly and widely applied in many organisms for delicate genetic manipulation, including human-induced pluripotent stem cells (iPSCs). Gene editing in human iPSCs is...
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Investigating Fungal Biosynthetic Pathways Using Pichia pastoris as a Heterologous Host
Fungal natural products have extensive biological activities, and thus have been largely commercialized in the pharmaceutical, agricultural, and food industries. Recently, heterologous expression has become an...
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Reactive Oxygen Species-Responsive and Self-Illuminating Nanoparticles for Inflammation and Tumor Imaging
Reactive oxygen species (ROS) play a key role in various physiological and pathological processes. Abnormally elevated ROS levels are generally related to the pathogenesis of inflammatory diseases and tumors. ...
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Gene Co-expression Network Analysis and Linking Modules to Phenoty** Response in Plants
Environmental factors, including different stresses, can have an impact on the expression of genes and subsequently the phenotype and development of plants. Since a large number of genes are involved in respon...
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Functional Analysis of Actin-Binding Proteins in the Central Nervous System of Drosophila
Using Drosophila actin-binding protein Dunc-115 as an example, this chapter describes a MARCM (mosaic analysis with a repressible cell marker)-based method for analyzing cytoskeletal components for their function...
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Integration of Omics and Phenotypic Data for Precision Medicine
Over the past two decades, biomedical research is moving toward a big-data-driven approach. The underlying causes of this transition include the ability to gather genetic or molecular profiles of humans faster...
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Single Transcript Unit CRISPR 2.0 Systems for Genome Editing in Rice
CRISPR-Cas9 and Cas12a (formerly Cpf1), RNA-guided DNA endonucleases found from adaptive immune system in prokaryotes, have been engineered and widely adopted as two of the most powerful genome editing systems...
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Improving a Quantitative Trait in Rice by Multigene Editing with CRISPR-Cas9
CRISPR-Cas9 system is one sequence-specific nuclease (SSN) that has several advantages over zinc finger nuclease (ZFN) and transcription activator-like effector nuclease (TALEN), such as multiplex genome editi...
