التفاصيل البيبلوغرافية
| العنوان: |
CRISPR Technique Incorporated with Single-Cell RNA Sequencing for Studying Hepatitis B Infection |
| المؤلفون: |
Connie Le (8463771), Yanming Liu (1482403), Joaquín López-Orozco (11210140), Michael A. Joyce (9694450), X. Chris Le (17762), D. Lorne Tyrrell (17808) |
| سنة النشر: |
2021 |
| المجموعة: |
Smithsonian Institution: Digital Repository |
| مصطلحات موضوعية: |
Biophysics, Microbiology, Cell Biology, Genetics, Molecular Biology, Immunology, Developmental Biology, Cancer, Infectious Diseases, Virology, Biological Sciences not elsewhere classified, Chemical Sciences not elsewhere classified, CRISPR Technique Incorporated, liver cell model, CRISPR-Cas 9 technique, hepatitis B virus, HBV infection, HBV RNA, HBV-infected Huh 7.5-NTCP cells, Huh 7.5-NTCP cells, Hepatitis B Infection, gene expression, CRISPR-mediated enrichment technique, Single-Cell RNA Sequencing, cell RNA-seq analysis, transcript |
| الوصف: |
Single-cell RNA sequencing (scRNA-seq) provides rich transcriptomic information for studying molecular events and cell heterogeneity at the single-cell level. However, it is challenging to obtain sequence information from rare or low-abundance genes in the presence of other highly abundant genes. We report here a CRISPR-Cas9 technique for the depletion of high-abundance transcripts, resulting in preferential enrichment of rare transcripts. We demonstrate an application of this CRISPR-mediated enrichment technique to scRNA-seq of liver cells infected with hepatitis B virus (HBV). Direct sequencing without the CRISPR-mediated enrichment detected HBV RNA in only 0.6% of the cells. The CRISPR-mediated depletion of the three most abundant transcripts resulted in selective enrichment of the HBV transcript and successful sequencing of HBV RNA in more than 74% of the cells. The improvement enabled a study of HBV infection and interferon treatment of a liver cell model. Gene clusters between the control and HBV-infected Huh7.5-NTCP cells were similar, suggesting that HBV infection did not significantly alter gene expression of the host cells. The treatment with interferon alpha dramatically changed the gene expression of Huh7.5-NTCP cells. These results from the single cell RNA-seq analysis of 7370 cells are consistent with those of bulk experiments, suggesting that HBV is a “stealth virus”. |
| نوع الوثيقة: |
article in journal/newspaper |
| اللغة: |
unknown |
| العلاقة: |
https://figshare.com/articles/journal_contribution/CRISPR_Technique_Incorporated_with_Single-Cell_RNA_Sequencing_for_Studying_Hepatitis_B_Infection/15082750Test |
| DOI: |
10.1021/acs.analchem.1c02227.s001 |
| الإتاحة: |
https://doi.org/10.1021/acs.analchem.1c02227.s001Test |
| حقوق: |
CC BY-NC 4.0 |
| رقم الانضمام: |
edsbas.1C69CC6B |
| قاعدة البيانات: |
BASE |
