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RNA Seq Experiment. The single-cell RNA-seq technique converts a population of RNAs to a library of cDNA fragments. These fragments are sequenced by high-throughput next generation sequencing techniques and the reads are mapped back to the reference genome, providing a count of the number of reads associated with each gene. [13]
Single-cell RNA sequencing (scRNA-seq) provides the expression profiles of individual cells and is considered the gold standard for defining cell states and phenotypes as of 2020. [44] Although it is impossible to obtain complete information on every RNA expressed by each cell, due to the small amount of material available, gene expression ...
RNA-Seq can also be used to determine exon/intron boundaries and verify or amend previously annotated 5' and 3' gene boundaries. Recent advances in RNA-Seq include single cell sequencing, bulk RNA sequencing, [6] 3' mRNA-sequencing, in situ sequencing of fixed tissue, and native RNA molecule sequencing with single-molecule real-time sequencing. [7]
Emerging technologies are aiming to combine pooled CRISPR screens with the detailed resolution of massively parallel single-cell RNA-sequencing (RNA-seq). Studies utilising “CRISP-seq”, [ 75 ] “CROP-seq”, [ 76 ] and “PERTURB-seq” [ 77 ] have demonstrated rich genomic readouts, accurately identifying gene expression signatures for ...
Perturb-seq is a reverse genetics approach that allows for the investigation of phenotypes at the level of the transcriptome, to elucidate gene functions in many cells, in a massively parallel fashion. The Perturb-seq protocol uses CRISPR technology to inactivate specific genes and DNA barcoding of each guide RNA to allow for all perturbations ...
Single-cell RNA sequencing (scRNA-seq) is a recently developed technique that allows the analysis of the transcriptome of single cells, including bacteria. [25] With single-cell transcriptomics, subpopulations of cell types that constitute the tissue of interest are also taken into consideration. [26]
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