CD8-positive, alpha-beta memory T cell — IGVFSM5367RVVU

released
Summary
CD8-positive, alpha-beta memory T cell, Homo sapiens modified with CRISPRko Sp-Cas9, transduced (lentivirus) with a guide library (MOI of 0.4)
Sample Terms
Sex
unspecified
Age
unknown
Taxa
Homo sapiens
Description
Lentivirus encoding for ZNF217 targeting gRNA
Sources
Submitter Comment
CD8+ T cells were activated with CD3/CD28 dynabeads at a 1:1 ratio. At 24 hours post-activation, CD8+ T cells were transduced with gRNA lentivirus encoding a ZNF217 targeting gRNA. At 48 hours post-activation, 3.5 x 10^5 T cells were electroporated with Cas9 protein. Briefly, cells were collected, spun down at 90xg for 10 minutes, resuspended in 20uL of Lonza P3 Primary buffer with 1.12 ug Cas9, and electroporated with the pulse code EH115. Cells were cultured in PRIME-XV T cell expansion XSFM (FujiFilm) supplemented with 5% human platelet lysate (Compass Biomed), 100 units/mL of penicilllin, 100 units/mL of streptomycin, and 100 units/mL human IL-2 (Peprotech). T cells were maintained at a concentration of 1-2e6 per mL for 9 days before RNA-sequencing. RNA was harvested using Norgen's Total RNA Purification Kit. RNA libraries for RNA-seq were prepared by Azenta Life Sciences.
Aliases
charles-gersbach:cd8-tcells-primary-cell-cas9-ko-znf217-rep3
Publications
Institutional Certificates

Donors

1 item
Accession
Aliases
Sex
Taxa
  • charles-gersbach:cd8-tcells-donor-14
unspecified
Homo sapiens

File Sets

4 items
CRISPR knockout RNA-seq integrating a guide (sgRNA) library targeting transcription start sites in ZNF217
  • charles-gersbach:cd8-tcells-rna-seq-cas-ko-znf217-rep3
Charles Gersbach, Duke
Homo sapiens GRCh38 guide RNAs
  • charles-gersbach:cd8-tcells-primary-cell-cas9-ko-znf217-grnas-set
Charles Gersbach, Duke
knockout RNA-seq: transcriptome alignments
  • charles-gersbach:cd8-tcells-intermediate-analysis-rna-seq-cas-ko-znf217
Charles Gersbach, Duke
knockout RNA-seq: fold change over control, transcript quantifications
  • charles-gersbach:cd8-tcells-analysis-set-rna-seq-cas-ko-znf217
Charles Gersbach, Duke

Modifications

1 item
Summary
Modality
knockout

Attribution