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SRX23645633: GSM8083730: SOX2 knockdown bulk RNA-seq G.i1_clone2 +Dox biol_rep3; Gorilla gorilla; RNA-Seq
1 ILLUMINA (NextSeq 1000) run: 9.1M spots, 843.1M bases, 284.3Mb downloads

External Id: GSM8083730_r1
Submitted by: Ludwig-Maximilians-Universität München
Study: Generation and characterization of inducible KRAB-dCas9 iPSCs from primates for cross-species CRISPRi
show Abstracthide Abstract
Comparisons of molecular phenotypes across primates provide unique information to understand human biology and evolution and single-cell RNA-seq CRISPR interference screens are a powerful approach to analyze them. Here, we generate and validate three human, three gorilla and two cynomolgus iPS cell lines that carry a dox-inducible KRAB-dCas9 construct in the AAVS1 locus. We show that despite variable expression levels of KRAB-dCas9 among lines, comparable downregulation of target genes and comparable phenotypic effects are observed in a single-cell RNA-seq CRISPR interference screen. Hence, we provide valuable resources for performing and further extending CRISPRi screens in human and non-human primates. Overall design: Three human, three gorilla, and two cynolmolgus inducible KRAB-dCas9 iPS cell lines with an integrated SOX2-targeting gRNA were culture in +dox or -dox conditions for 4 days to compare between SOX2-knockdown (+dox) and normal condition (-dox).
Sample: SOX2 knockdown bulk RNA-seq G.i1_clone2 +Dox biol_rep3
SAMN39968303 • SRS20479782 • All experiments • All runs
Organism: Gorilla gorilla
Library:
Name: GSM8083730
Instrument: NextSeq 1000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Cells that were cultured in 24-wells, were lysed in 200 µL Buffer RLT Plus supplemented with 1 % 2-Mercaptoethanol and then stored at -80 °C until processing. Libraries were constructed following the published prime-seq protocol. (Janjic, A., Wange, L.E., Bagnoli, J.W. et al. Prime-seq, efficient and powerful bulk RNA sequencing. Genome Biol 23, 88 (2022). https://doi.org/10.1186/s13059-022-02660-8); https://www.protocols.io/view/prime-seqs9veh66 polyA-capture bulk RNA-seq
Runs: 1 run, 9.1M spots, 843.1M bases, 284.3Mb
Run# of Spots# of BasesSizePublished
SRR279928789,065,638843.1M284.3Mb2024-02-20

ID:
31925123

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