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| Status |
Public on Sep 13, 2022 |
| Title |
Loss of Zfp335 triggers cGAS/STING-dependent apoptosis of post-B-selection pre-T cells [TCRa-seq] |
| Organism |
Mus musculus |
| Experiment type |
Other
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| Summary |
The transcription factor Zfp335 is esstential for the survival of post-B-selection DN4 thymocytes. We utilized TCRa repertoire sequencing to assess alterations to survival duration for Zfp335-deficient DP thymocytes.
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| Overall design |
DP thymocytes were FACS-sorted (Live Lin- CD4+ CD8+) from Zfp335-fl/fl or Zfp335-fl/fl E8III-cre positive 7 week old male and female mice. Each sample was from one individual mouse. Following sorting, total RNA was isolated and libraries were prepared as follows: 5ng total RNA was reverse transcribed with Trac-specific reverse transcription primer and template switch oligo containing a Unique Molecular Identifier (UMI) using SmartScribe RT (Takara Bio). cDNA was purified and TCRa transcripts were amplified with a TSO-specific nested Trac-specific primers using Q5 polymerase (NEB). Each sample was dual indexed then all samples pooled and Illumina-compatible adaptors were added using the NEBNext Ultra II DNA library prep kit. 300x300bp sequencing was performed on an Illumina MiSeq instrument.
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| Contributor(s) |
Ratiu J |
| Citation(s) |
36202870 |
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| Submission date |
Jun 01, 2022 |
| Last update date |
Oct 27, 2022 |
| Contact name |
Jeremy Ratiu |
| E-mail(s) |
Jeremy.Ratiu@duke.edu
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| Phone |
5868505692
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| Organization name |
Duke University
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| Department |
Immunology
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| Lab |
Zhuang/ Shinohara
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| Street address |
328 Jones Bldg, Box 3010
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| City |
Durham |
| State/province |
North Carolina |
| ZIP/Postal code |
27710 |
| Country |
USA |
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| Platforms (1) |
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| Samples (7)
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| Relations |
| BioProject |
PRJNA844384 |
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