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Technique Concept

Sequence Verification

Confirming construct identity by Sanger or next-generation sequencing.

67 total methods

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Methods

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single-cell RNA sequencing

Single-cell RNA sequencing (scRNA-seq) is a transcriptomic assay method that measures RNA molecules in individual cells by sequencing-based transcript detection. In the cited application, it detected FLiCRE transcripts within the endogenous transcriptome, enabling simultaneous readout of cell type and calcium activation history.

Adeno-associated virus

Adeno-associated virus (AAV) is a viral delivery harness used to package and express CRISPR genome-editing components in vivo. In the cited literature, AAV supports single-vector delivery when smaller Cas9 orthologues and their chimeric guide RNAs fit within AAV packaging constraints, enabling robust in vivo genome editing.

RNA sequencing

RNA sequencing (RNA-seq) is a transcriptomic assay method that quantifies gene-expression changes by sequencing RNA-derived libraries. In the cited study, it was used on adult rat amygdala tissue to detect subtle expression changes associated with development, cellular function, and nervous system disease after gestational high-THC cannabis smoke exposure.

immunohistochemistry

Immunohistochemistry is an antibody-based tissue staining assay used in the cited stroke study alongside transcriptomics and real-time polymerase chain reaction to examine post-stroke tissue in aged rats and post-stroke patients. In that context, it supported assessment of angiogenesis-related histological features such as vascular density after stroke.

chromatin immunoprecipitation sequencing

Chromatin immunoprecipitation sequencing (ChIP-seq) is an assay method that combines chromatin immunoprecipitation with sequencing-based genomic localization to map protein-associated genomic regions. In the cited study, it was used to identify genome-wide ZFHX3-binding sites in suprachiasmatic nucleus chromatin, revealing occupancy concentrated near transcription start sites and co-localization with known histone modifications.

next-generation sequencing

Multi-locus sequencing, multiplex PCR, and next-generation sequencing refined the identification of fungal and bacterial pathogens.

chemogenetics

Chemogenetics is an engineering method in which target proteins are genetically engineered to interact with a designed chemical partner with high selectivity. It is used as a chemical-input strategy to manipulate protein or receptor function in cells and has also been used alongside optogenetics to perturb cellular structures such as specific microtubule subtypes.

spatial transcriptomics

Spatial transcriptomics is a transcriptomic assay method identified in the supplied review as a recent methodological advance. In that evidence, it is presented as part of a broader technology set that enables easier and more accurate visualization of cell behavior and qualitative and quantitative analysis of cell-cell interactions.

calcium imaging

These tools include quantitative behavioral assays, calcium imaging, optogenetics and transgenics.

single-cell sequencing

This review highlights the latest cutting-edge technologies driving progress in the field, including ... single-cell sequencing...

light-controlled crRNA

Light-controlled crRNA is a chemically modified CRISPR guide RNA in which vitamin E is attached to the 5' terminus through a photolabile linker, creating a caged crRNA that inactivates CRISPR/Cas9 until light exposure. Upon irradiation, this design restores CRISPR/Cas9 activity and supports genome editing of VEGFA and knockdown of EGFP expression in human cells.

Ptac-lacIq regulatory system

The pUC18T-mini-Tn7T system enables IPTG-inducible, site-specific genomic integration of genes in single copy via the Ptac-lacIq regulatory system.

bisulfite pyrosequencing

Bisulfite pyrosequencing is an assay method used to quantify promoter CpG DNA methylation after bisulfite treatment. In the cited study, it was used to measure methylation in the CAMK1D, CRY2, and CALM2 promoter regions in peripheral blood from patients with type 2 diabetes and matched healthy controls.

H3K36me3 cell-free chromatin immunoprecipitation sequencing

H3K36me3 cell-free chromatin immunoprecipitation sequencing (cfChIP-seq) is a plasma-based assay that establishes a personal gene expression profile from cell-free chromatin. In the cited study context, it functions as a reference enrichment assay for active genes in liquid biopsy samples.

proximity labeling

Proximity labeling is described here as a methodological approach proposed to define state-specific proteomic and post-translational signatures in studies evaluating the addivosome pathological condensate model. The supplied evidence does not identify a specific proximity-labeling enzyme, chemistry, or construct design.

SOSHI-seq

We present a cheap, rapid, and versatile assay called SOSHI-seq (Screening of Self-transcribed Hormone Inducible response elements coupled to sequencing).

next-generation sequencing

Sensitive detection of tobamoviruses in the field with minimal sample preparation can be achieved using latest technologies such as isothermal amplification, CRISPR/Cas-hybrid assays or next-generation sequencing.

multiparametric flow cytometry

advanced tools such as next-generation sequencing (NGS), direct stochastic optical reconstruction microscopy (dSTORM), or multiparametric flow cytometry are helping to identify novel tumor-specific targets and improve therapy designs

multiplex PCR

Multi-locus sequencing, multiplex PCR, and next-generation sequencing refined the identification of fungal and bacterial pathogens.

single unique molecular identifier strategy

Here, we implement a single unique molecular identifier strategy that reduces sequencing artifacts and achieves an error rate of ~10⁻⁵, enabling single-particle-level quantification of quasi-species diversity.

vesicle fluid culture

Both vesicle fluid culture and next-generation sequencing (NGS) of blood samples confirmed the presence of Escherichia coli.

dual enzyme lineage tracing system

The dual enzyme lineage tracing system is identified in a 2023 review as a recent methodological advance in lineage tracing. The supplied evidence indicates that it is intended to support visualization of cell behavior and analysis of cell-cell interactions, but no construct-level, enzyme-specific, or operational details are provided.

Immunoblot

By using soy-absorbed, peanut-allergic patient sera on two-dimensional immunoblots and N-terminal amino-acid sequencing, about 30 protein fractions were shown to be isoforms, or fractions, of the three major peanut proteins.

polymerase chain reactions

While these methods, like polymerase chain reactions or whole-genome sequencing, are considered the "gold standard" for diagnostics, the development of inexpensive, rapid diagnostic assays is necessary for effective AMR detection and management.

programmable genetic circuits

Programmable genetic circuits are engineered genetic constructs used to create designer cells with controllable behaviors in mammalian synthetic biology. The cited literature describes circuits that can incorporate targetable DNA-binding systems such as CRISPR/Cas9 and sensor-actuator devices to regulate complex cellular functions with high spatial and temporal resolution.

sequencing-based solutions

Sequencing-based solutions are proposed assay methods for detecting large-scale CRISPR-associated genomic alterations. In the cited review, they are positioned as potential approaches to identify rare events such as translocations, inversions, deletions, and chromothripsis that can be missed by current workflows.

mRNA-sequencing

mRNA-sequencing of OX lines revealed substantial upregulation of Pi starvation-responsive genes, including SPX1, PS2/PECP2, and SRG3/GDPD1.

small RNA-sequencing

small RNA-sequencing identified the downregulation of miR399 and miR827, which are involved in the Pi starvation response.

CRISPR/Cas-hybrid assays

Sensitive detection of tobamoviruses in the field with minimal sample preparation can be achieved using latest technologies such as isothermal amplification, CRISPR/Cas-hybrid assays or next-generation sequencing.

pooled library approach

The pooled library approach is an engineering method for rapid generation and parallel screening of nearly all possible split-protein constructs, with sequencing-based readout. In the cited application, it was used with optogenetic dimers to comprehensively map split-site behavior across Cre recombinase and support inducible post-translational control design.

MeRIP-seq

high-throughput technologies such as MeRIP-seq and nanopore direct RNA sequencing have enabled the preliminary construction of RNA methylation landscapes in cucurbit species

nanopore direct RNA sequencing

high-throughput technologies such as MeRIP-seq and nanopore direct RNA sequencing have enabled the preliminary construction of RNA methylation landscapes in cucurbit species

imaging

this view has been challenged in the last few years with the availability of RNA sequencing, immunohistochemistry, electron microscopy, morphological reconstruction, and imaging data

morphological reconstruction

this view has been challenged in the last few years with the availability of RNA sequencing, immunohistochemistry, electron microscopy, morphological reconstruction, and imaging data

spatial proteomics

The emerging tools that have translational relevance, as pointed out by the review, include... spatial proteomics...

synaptic positron emission tomography imaging

The emerging tools that have translational relevance, as pointed out by the review, include... synaptic positron emission tomography (PET) imaging, with the capabilities of providing disease-specific and patient-level insights into the pathology of synapses.

whole exome sequencing

Whole exome sequencing identified two heterozygous variants in the NPHS1 gene, as well as one heterozygous and one homozygous variant in the PLEKHG2 gene.

electron microscopy

Light and electron microscopy have been widely used in confirming the bipolarity of somatic embryos during somatic embryogenesis.

long non-coding RNA

Long non-coding RNAs (lncRNAs) are non-protein-coding RNA elements within the broader non-coding RNA landscape. In plants, available reports indicate that lncRNAs participate in light-dependent processes including photomorphogenesis, cotyledon greening, and photoperiod-regulated flowering.

rs1800378-targeted allele-selective CRISPR/Cas9 VWF disruption strategy

Our strategy permanently inactivates VWF variants by selectively disrupting the pathogenic allele's open reading frame via the introduction of indels by Cas9. To circumvent the challenge of designing variant-specific strategies, we targeted the common single nucleotide polymorphism (SNP) rs1800378 in VWF.

multiomics sequencing

The review further evaluates the clinical translational potential and challenges of innovative strategies such as vagus nerve stimulation, optogenetics, multiomics sequencing, and cytokine-targeted therapies.

synthetic receptor system

The synthetic receptor system is described in a 2023 review as a methodological advance for studying cell behavior and cell-cell interactions. The available evidence indicates that it enables easier and more accurate visualization and supports qualitative and quantitative analysis in this context, but no molecular architecture or operating principle is provided.

CellChat

We then develop CellChat, a tool that is able to quantitatively infer and analyze intercellular communication networks from single-cell RNA-sequencing (scRNA-seq) data.

CNNeoPP

Here, we present CNNeoPP, an integrated computational pipeline for neoantigen discovery. CNNeoPP was rigorously validated using independent datasets, including the TESLA dataset, and experimental validation via ELISpot T-cell assays.

CRISPR screening

Personalization using advanced technologies like CRISPR screening and single-cell RNA sequencing can enhance durability and effectiveness of treatments for heavily pretreated patients

Dogme

we developed Dogme to automate basecalling, alignment, modification detection, and transcript quantification. Dogme automates the reprocessing of ONT POD5 files

FragmentFinder

we have developed FragmentFinder-an intuitive, Windows-executable resource designed to require absolutely no computational background and capable of accurately characterizing all (annotated and unknown) sncRNA-derived RNAs within a raw small RNA sequencing file in real time.

gp64 variant calling methodology

we describe a variant calling methodology that allows the detection of the targeted mutations in gp64 even though these mutations are not the dominant sequences... our sequencing pipeline successfully detected indel mutations within gp64 for most of the single-guide RNA (sgRNA) targets

iFish

Finally, we established the iFish database (https://gonglab.hzau.edu.cn/iFish/), a user-friendly platform that supports interactive browsing, visualization, and download.

incoherent feed-forward loop

We perform RNA-sequencing on cells expressing an incoherent feed-forward loop (iFFL), a genetic circuit that enhances operational capacity.

light-driven ion pump rhodopsins

Ion-transporting microbial rhodopsins are widely used as major molecular tools in optogenetics... light-driven ion pumps actively transport specific ions, such as H+, Na+, Cl-, against electrophysiological potential by using light energy... light-driven ion-pumping rhodopsins are mostly applied as inhibitory optogenetics tools.

LnCeVar 2.0

LnCeVar 2.0 is an updated database investigating genomic variations that disrupt competing endogenous RNA (ceRNA) networks via single-cell and spatial transcriptomics.

miniscopes

The ability to simultaneously image the spatiotemporal activity signatures from many neurons during unrestrained vertebrate behaviors has become possible through the development of miniaturized fluorescence microscopes, or miniscopes, sufficiently light to be carried by small animals such as bats, birds and rodents.

NGSTroubleFinder

Here, we present NGSTroubleFinder, a novel tool to detect cross-sample contamination in human whole-genome and whole-transcriptome sequencing data, sample swaps, and mismatches between the reported and the inferred genetic and transcriptomic sexes.

PB-TRIBE-STAMP

we developed PB-TRIBE-STAMP, a tool based on two orthogonal RNA editing enzymes

RMPore

We developed RMPore (https://rmpore.renlab.cn/), a comprehensive database of single-molecule RNA modifications detected from 958 DRS samples across 34 species.

SCAN-ACT

We developed a comprehensive computational pipeline called SCAN-ACT that leverages single-cell RNA sequencing and multi-omics data from tumor and normal tissues to nominate and prioritize putative targets for both chimeric antigen receptor (CAR)- and TCR-T cells.

scSNViz

scSNViz is an R package for the exploration, quantification, and visualization of expressed single-nucleotide variants (SNVs) from cell-barcoded single-cell RNA sequencing (scRNA-seq) data.

single-cell RNA-seq

The supplied web research summary states that the anchor review emphasizes mechanistic advances enabled by optogenetics, RNA-sequencing, animal models, and human genetics, and specifically highlights nociceptor and somatosensory neuron classification by single-cell RNA-seq.

single-nucleus RNA sequencing

Recent advances in spatial transcriptomics, single-nucleus RNA sequencing, chemogenetics, and optogenetics are discussed as transformative tools for mapping and manipulating VOR-expressing circuits.

The Cancer Genome Atlas

The Cancer Genome Atlas (TCGA) is a public funded project that aims to catalogue and discover major cancer-causing genomic alterations to create a comprehensive "atlas" of cancer genomic profiles.

TSCS

The web research summary states that the anchor paper describes TSCS (Transcript SNVs Classifier relying on complementary sequencings), a machine-learning framework that integrates short-read MGI RNA-seq with long-read PacBio RNA-seq to improve transcript SNV discovery and to distinguish genomic transcript SNVs from RNA-editing-derived transcript SNVs.

whole-genome sequencing

While these methods, like polymerase chain reactions or whole-genome sequencing, are considered the "gold standard" for diagnostics, the development of inexpensive, rapid diagnostic assays is necessary for effective AMR detection and management.