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| Status |
Public on Jun 23, 2020 |
| Title |
Gene expression of Abhd11 knock-out or over-expression embryonic stem cells |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Abhd11 is previously identified as a conserved lipase in lipid metabolism. However, its physiological functions in embryonic stem cells (ESCs) are unknown. Based on the construction of ESC lines with conditional Abhd11 expression, we analyst the transcriptome profiling of ESCs upon Abhd11 knock-out (KO) or over-expression (OE). In Abhd11 KO ESCs, differentially expressed genes were enriched in metabolic genes while Abhd11 OE ESCs elicited a smaller transcriptional response compared to that of Abhd11 KO ESCs.
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| Overall design |
Abhd11 was deleted under the inducible overexpression (iOE) of exogenous Abhd11 using “tetR-Krab-tetO” conditional gene expression system and CRISPR/Cas9 technology. To start with, we generated a stable tetR-Krab-Cas9 expressing cell line, which was hereafter used as a control cell lines. Next, we obtained Abhd11 iOE ESCs by transducing inducible expression cassette of Abhd11 targeted with Red fluorescent protein (Abhd11-RFP) into the tetR-Krab-Cas9 expressing cells. The subcloned Abhd11 iOE ESCs was induced to express Abhd11-RFP and used as Abhd11 OE ESCs in the presence of 1000ng/ml doxycycline (Dox). Finally, we generated Abhd11 iOE/KO ESCs by transducing lentivirus carrying sgRNAs that mapped to the Abhd11 genomic sequence spanning an intron-exon junction and only targeted endogenous Abhd11 into the Abhd11 iOE ESCs. Hence, endogenous Abhd11 depleted Abhd11 iOE/KO ESCs were used as Abhd11 KO ESCs in the absence of Dox while used as Abhd11 rescued (Abhd11 RES) ESCs in the presence of 1ng/ml Dox.
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| Contributor(s) |
Liu G, Jian R |
| Citation(s) |
32733886 |
| Submission date |
Dec 16, 2019 |
| Last update date |
Aug 03, 2020 |
| Contact name |
Rui Jian |
| E-mail(s) |
jianruilq2@aliyun.com
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| Phone |
+8613627696789
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| Organization name |
Third Military Medical University
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| Department |
Department of Histology and Embryology
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| Lab |
Laboratory of Stem Cell and Developmental Biology
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| Street address |
Gaotanyan Street
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| City |
Chongqing |
| ZIP/Postal code |
400038 |
| Country |
China |
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| Platforms (1) |
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| Samples (10)
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| Relations |
| BioProject |
PRJNA595847 |
| SRA |
SRP237744 |
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