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cDNA Synthesis & Library Construction

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SMARTer ICELL8 Citations

Our collaborators and customers are constantly making scientific breakthroughs. Here are the latest published results obtained using the SMARTer ICELL8 Single-Cell System.

  1. Aarts, M. et al. Coupling shRNA screens with single-cell RNA-seq identifies a dual role for mTOR in reprogramming-induced senescence. Genes Dev. 31, 2085-2098 (2017).

    By carrying out single-cell RNA-seq and shRNA screening in parallel, the speed at which gene candidates from such functional screenings can be identified and validated can be significantly increased. The authors of this study investigated mediators of reprogramming-induced senescence. After an initial shRNA screen identified four candidate genes (MTOR, CDKN1A, MYOT, and UBE2E1) whose knockdown bypassed reprogramming-induced senescence, the SMARTer ICELL8 Single-Cell System was used to prepare RNA-Seq libraries on cells infected with shRNAs targeting each of these candidates. Further investigation revealed that mTOR inhibition has an antagonistic effect during cellular reprogramming. The SMARTer ICELL8 platform's flexible cell-size isolation and its available sensitive library preparation chemistries were key to accelarating discovery from the screening process by simultaneously allowing shRNA identification and transcriptome analysis from the RNA-seq data of the processed single cells.

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  2. Gao, R. et al. Nanogrid single-nucleus RNA sequencing reveals phenotypic diversity in breast cancer. Nat. Commun. 8, 228 (2017).

    The researchers utilized the SMARTer ICELL8 Single-Cell System to demonstrate strong concordance between transcriptional profiling data generated from single cells and nuclei obtained from a cancer cell line. Common single-cell RNA-seq protocols for transcriptome analysis are incompatible with cancer cells derived from flash frozen archival tissue specimens because the preservation process disrupts the cell membranes. The SMARTer ICELL8 platforms's flexibility to allow isolation of nuclei and fixed frozen cells enabled the generation of representative, high-quality RNA-seq data from the samples. This method provides a powerful new solution for working with archival samples because nuclear membranes are not usually disrupted by freeze-thaw cycles. In the article, the authors state that "transcriptome profiles of nuclei are highly representative of whole cells, and can be used to study many cancer genes and signaling pathways."

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  3. Goldstein, L. D. et al. Massively parallel nanowell-based single-cell gene expression profiling. BMC Genomics 18, 519 (2017).

    In this study, RNA-seq was performed to profile the transcriptomes of single cells derived from cultured cells and complex tissues. Using the SMARTer ICELL8 Single-Cell System, the researchers were able to discriminate profiles obtained from a mixture of human and mouse cells processed on a single chip, as demonstrated by a low multiplet rate (<3%). Furthermore, minimal cross-contamination was observed in this experiment as indicated by the high single-cell purity (94-97%). Other findings show the ability to distinguish representative cell types within mouse pancreatic islet samples. The SMARTer ICELL8 platform's imaging capabilities for identifying single-cell-containing wells from empty and multiple-cell-containing wells was crucial for ensuring the high single-cell purity and low multiplet rate observed in this study.

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