|
| 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
|
| 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.
|
| |
|
| 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.
|
| |
|
| Contributor(s) |
Ratiu J |
| Citation(s) |
36202870 |
| |
| Submission date |
Jun 01, 2022 |
| Last update date |
Oct 27, 2022 |
| Contact name |
Jeremy Ratiu |
| E-mail(s) |
[email protected]
|
| Phone |
5868505692
|
| Organization name |
Duke University
|
| Department |
Immunology
|
| Lab |
Zhuang/ Shinohara
|
| Street address |
328 Jones Bldg, Box 3010
|
| City |
Durham |
| State/province |
North Carolina |
| ZIP/Postal code |
27710 |
| Country |
USA |
| |
|
| Platforms (1) |
|
| Samples (7)
|
|
| Relations |
| BioProject |
PRJNA844384 |
Less...
More...