|
| Status |
Public on Feb 28, 2023 |
| Title |
JB82 THC/SIV |
| Sample type |
RNA |
| |
|
| Source name |
Gingiva
|
| Organism |
Macaca mulatta |
| Characteristics |
(treatment): SIV infected and THC treated
|
| Treatment protocol |
10 animals were infected intravenously with 100 TCID50 SIVmac251 and 4 remained uninfected. Six SIV-infected rhesus macaques were administered delta-9-tetrahydrocannabinol at 0.18 mg/kg one month before SIV Infection and increased to 0.32 mg/kg on the day of SIV infection until 6 months post SIV infection. Four SIV-infected rhesus macaques were administered the same volume of vehicle one month before SIV Infectionuntil 6 months post SIV infection.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA (small and large) was extracted using the miRNeasy Mini kit (Qiagen). On-column DNase digestion was also performed to eliminate contaminating genomic DNA.
|
| Label |
n/a
|
| Label protocol |
microRNA profiling was performed using the TaqMan® OpenArray® Human microRNA panels (Life Tech) following the manufacturer’s instructions. Reverse transcription was first performed from 100 ng of total RNA using the microRNA reverse transcription kit. 2.5 µL of cDNA was preamplified using the TaqMan® PreAmp Master Mix kit. Preamplified products were first diluted and then loaded onto to OpenArray plates. OpenArray plates were run on the QuanStudio™ 12K Flex Real-Time PCR system (ThermoFisher Scientific) following the manufacturer’s recommended thermal cycling conditions.
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
Test-2
|
| Data processing |
QuantStudio run files representing all treatment groups and timepoints were analyzed using ExpressionSuite Software (Life Tech). The results from the ExpressionSuite software analysis containing five columns (well, sample, detector, task and CT values) were saved as a tab-delimited text file which was later imported and analyzed using the Omics Office StatMiner qPCR analysis software, TIBCO Spotfire, (Perkin Elmer, Waltham, MA). Omics Office StatMiner Software utilizes the comparative Cτ (ΔΔCτ) method to rapidly and accurately quantify relative gene expression across many genes and samples.
miRNA expression data was normalized using the global normalization method.
In all experiments, the CT upper limit was set to 28 meaning that all miRNA detectors with a CT value greater than or equal to 28 were excluded.
Due to the small sample size and exploratory nature of the study multiple comparisons correction was not applied. However, differentially expressed miRNAs were confirmed using RT-qPCR.
Omics Office StatMiner qPCR analysis software automatically calculates all deltadeltaCT based fold-change calculations and provides p values from the uploaded QuantStudio data files.
Matrix normalized worksheet reports normalized signal (Normalization using global normalization method).
Fold Change worksheet reports test-1/control (i.e., VEH/SIV vs Uninfected control) and test-1 vs test-2 (THC/SIV vs VEH/SIV) ratios.
|
| |
|
| Submission date |
Dec 12, 2022 |
| Last update date |
Feb 28, 2023 |
| Contact name |
Mahesh Mohan |
| E-mail(s) |
mmohan@txbiomed.org
|
| Organization name |
Southwest National Primate Research Center
|
| Department |
Host Pathogen Interaction Program
|
| Lab |
12/106
|
| Street address |
8715 West Military Road
|
| City |
San Antonio |
| State/province |
Texas |
| ZIP/Postal code |
78227 |
| Country |
USA |
| |
|
| Platform ID |
GPL17837 |
| Series (2) |
| GSE220709 |
MicroRNA profiling of Basal ganglia tissue from chronically SIV infected rhesus macaques II |
| GSE220783 |
Cannabinoid modulation of gene and microRNA expression in brain (basal ganglia) during chronic HIV/SIV infection |
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