1. Uporaba transkriptomike na nivoju posamezne celice
- Author
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Škoda, Neža and Jakše, Jernej
- Subjects
Illumina ,mikropretočni sistemi ,applications of transcriptomics ,microfluidics ,transkriptomika posamezne celice ,single cell transcriptomics ,uporaba transkriptomike - Abstract
Transkriptom so vse molekule RNA v celici ali tkivu. Transkriptomika je veda, ki preučuje transkriptom in pomaga pri razumevanju regulacije izražanja genov in bioloških procesov. Sekvenciranje transkriptoma iz tkiv pokaže izražanje genov velikega števila celic. Transkriptomika na nivoju posamezne celice pa omogoča vpogled v gensko izražanje ene celice. Ponuja boljši prikaz razlik med posameznimi celicami in njihovimi funkcijami. Da se lahko spustimo na raven ene celice moramo celice najprej ločiti med sabo. Preučujemo lahko specifične redke celične tipe ali pa obdelamo veliko število celic naenkrat. Celice lahko ločujemo tudi po velikosti in s pomočjo fluorescence, glede na njihove molekularne markerje. Prepise pridobljenih posameznih celic unikatno označimo in pripravimo knjižnico za sekvenciranje. Sekvenciramo z visokozmogljivimi tehnologijami, ki delujejo na principu sekvenciranja s sintezo. Pridobljene podatke nato računalniško obdelamo. Obstaja tudi metoda sekvenciranja transkriptoma posameznih jeder, ki omogoča raziskovanje celic, ki jih je težko izolirati iz tkiva in zamrznjenih vzorcev. Metoda preučevanja transkriptoma na nivoju ene celice se uveljavlja na številnih področjih. V raziskavah zgodnjega embrionalnega razvoja omogoča vpogled v uravnavanje izražanja genov že v fazi morule. V nevroznanosti omogoča spremljanje diferenciacije celic. Pri razvoju tumorjev je omogočila iskanje novih tarč za zdravila pri redkih populacijah celic. Uveljavlja se v raziskavah rastlin. The transcriptome is the set of RNA molecules in a cell or tissue. Transcriptomics is the study of the transcriptome to understand the regulation of gene expression and biological processes. Sequencing the transcriptome of tissues shows gene expression in many cells. Transcriptomics at the single cell level provides insight into the gene expression of a single cell. This gives a better indication of the differences between individual cells and their different functions. To get down to the level of a single cell, we need to first separate the cells. You can search for specific rare cell types or process many cells at once. Cells can also be distinguished by their size and fluorescence, depending on their molecular markers. Once we have the individual cells, we uniquely label their transcripts and prepare the library for sequencing. We sequence using high-throughput technologies that work on the principle of sequencing by synthesis. The data are then processed by computer. A method for sequencing the transcriptome of individual cell nuclei has also been developed, allowing the study of cells that are difficult to extract from tissue and frozen samples. Transcriptomics at the single cell level is gaining traction in many fields. In studies of early embryonic development, it provides insights into the regulation of gene expression, even at the morula stage. In neuroscience, it enables monitoring of cell differentiation. In tumor development, it enabled the search for new drug targets in rare cell populations. It is used in plant research.
- Published
- 2022