Your browser doesn't support javascript.
loading
Hydrop enables droplet-based single-cell ATAC-seq and single-cell RNA-seq using dissolvable hydrogel beads.
De Rop, Florian V; Ismail, Joy N; Bravo González-Blas, Carmen; Hulselmans, Gert J; Flerin, Christopher Campbell; Janssens, Jasper; Theunis, Koen; Christiaens, Valerie M; Wouters, Jasper; Marcassa, Gabriele; de Wit, Joris; Poovathingal, Suresh; Aerts, Stein.
Afiliação
  • De Rop FV; VIB-KU Leuven/VIB Center for Brain & Disease Research, Leuven, Belgium.
  • Ismail JN; Laboratory of Computational Biology, Department of Human Genetics, KU Leuven, Leuven, Belgium.
  • Bravo González-Blas C; VIB-KU Leuven/VIB Center for Brain & Disease Research, Leuven, Belgium.
  • Hulselmans GJ; Laboratory of Computational Biology, Department of Human Genetics, KU Leuven, Leuven, Belgium.
  • Flerin CC; VIB-KU Leuven/VIB Center for Brain & Disease Research, Leuven, Belgium.
  • Janssens J; Laboratory of Computational Biology, Department of Human Genetics, KU Leuven, Leuven, Belgium.
  • Theunis K; VIB-KU Leuven/VIB Center for Brain & Disease Research, Leuven, Belgium.
  • Christiaens VM; Laboratory of Computational Biology, Department of Human Genetics, KU Leuven, Leuven, Belgium.
  • Wouters J; VIB-KU Leuven/VIB Center for Brain & Disease Research, Leuven, Belgium.
  • Marcassa G; Laboratory of Computational Biology, Department of Human Genetics, KU Leuven, Leuven, Belgium.
  • de Wit J; Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, United States.
  • Poovathingal S; VIB-KU Leuven/VIB Center for Brain & Disease Research, Leuven, Belgium.
  • Aerts S; Laboratory of Computational Biology, Department of Human Genetics, KU Leuven, Leuven, Belgium.
Elife ; 112022 02 23.
Article em En | MEDLINE | ID: mdl-35195064
Scientists are now able to determine the order of chemical blocks, or nucleic acids, that make up the genetic code. These sequencing tools can be used to identify which genes are active within a biological sample. They do this by extracting and analysing open chromatin (regions of DNA that are accessible to the cell's machinery), or sequences of RNA (the molecular templates cells use to translate genes into working proteins). Initially, most sequencing tools could only provide an 'averaged-out' profile of the genes activated in bulk pieces of tissue which contain multiple types of cell. However, advances in technology have led to new methods that can extract and analyse open chromatin or RNA from individual cells. First, the cells are separated, via a technique called microfluidics, into tiny droplets of water along with a single bead that carries a unique barcode. The cell is then broken apart inside the droplet and the barcode within the bead gets released and attaches itself to the genetic material extracted from the cell. All the genetic material inside the droplets is then pooled together and sequenced. Researchers then use the barcode tags to identify which bits of RNA or DNA belong to each cell. Single-cell sequencing has many advantages, including being able to pinpoint precise genetic differences between healthy and abnormal cells, and to create cell atlases of whole organisms, tissues and microbial communities. But existing methods for extracting chromatin are very expensive, and there were no openly available tools for processing thousands of cells at speed. Furthermore, while several single-cell RNA sequencing tools are already freely available, they are not very sensitive or practical to use. Here, De Rop et al. have developed a new open-source platform called HyDrop that overcomes these barriers. The method entails a new type of barcoded bead and optimised elements of existing microfluidics protocols using open-source reagents. These changes created a more user-friendly workflow and increased sensitivity of sequencing at no additional cost. De Rop et al. used their new platform to screen the RNA and open chromatin of thousands of individuals cells from the brains of mice and flies. HyDrop outperformed other open-source methods when working in RNA-sequencing mode. It also provides the first open-source tool for sequencing open chromatin in single cells. Further improvements are expected as researchers tweak the platform, which for now provides an affordable alternative to existing methods.
Assuntos
Palavras-chave

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Sequenciamento de Nucleotídeos em Larga Escala / Sequenciamento de Cromatina por Imunoprecipitação Tipo de estudo: Guideline Limite: Animals Idioma: En Revista: Elife Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Bélgica País de publicação: Reino Unido

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Sequenciamento de Nucleotídeos em Larga Escala / Sequenciamento de Cromatina por Imunoprecipitação Tipo de estudo: Guideline Limite: Animals Idioma: En Revista: Elife Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Bélgica País de publicação: Reino Unido