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https://doi.org/10.1021/acssynbio.0c00499
Title: | Evaluating Capture Sequence Performance for Single-Cell CRISPR Activation Experiments | Authors: | Choo, Xin Yi Lim, Yu Ming Katwadi, Khairunnisa Yap, Lynn Tryggvason, Karl Sun, Alfred Xuyang Li, Shang Handoko, Lusy Ouyang, John F Rackham, Owen JL |
Keywords: | CRISPR activation hESC Overexpression Transcription Factors Single-cell RNAseq Feature barcoding technology |
Issue Date: | 19-Mar-2021 | Publisher: | AMER CHEMICAL SOC | Citation: | Choo, Xin Yi, Lim, Yu Ming, Katwadi, Khairunnisa, Yap, Lynn, Tryggvason, Karl, Sun, Alfred Xuyang, Li, Shang, Handoko, Lusy, Ouyang, John F, Rackham, Owen JL (2021-03-19). Evaluating Capture Sequence Performance for Single-Cell CRISPR Activation Experiments. ACS SYNTHETIC BIOLOGY 10 (3) : 640-645. ScholarBank@NUS Repository. https://doi.org/10.1021/acssynbio.0c00499 | Abstract: | The combination of single-cell RNA sequencing with CRISPR inhibition/activation provides a high-throughput approach to simultaneously study the effects of hundreds if not thousands of gene perturbations in a single experiment. One recent development in CRISPR-based single-cell techniques introduces a feature barcoding technology that allows for the simultaneous capture of mRNA and guide RNA (gRNA) from the same cell. This is achieved by introducing a capture sequence, whose complement can be incorporated into each gRNA and that can be used to amplify these features prior to sequencing. However, because the technology is in its infancy, there is little information available on how such experimental parameters can be optimized. To overcome this, we varied the capture sequence, capture sequence position, and gRNA backbone to identify an optimal gRNA scaffold for CRISPR activation gene perturbation studies. We provide a report on our screening approach along with our observations and recommendations for future use. | Source Title: | ACS SYNTHETIC BIOLOGY | URI: | https://scholarbank.nus.edu.sg/handle/10635/191306 | ISSN: | 21615063 | DOI: | 10.1021/acssynbio.0c00499 |
Appears in Collections: | Staff Publications Elements |
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Choo et al Manuscript.pdf | Submitted version | 2.99 MB | Adobe PDF | OPEN | Post-print | View/Download |
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