A single-cell CRISPRi platform for characterizing candidate genes relevant to metabolic disorders in human adipocytes.
CRISPRi
SGBS
adipocyte
adipogenesis
single-cell RNA-Seq
Journal
American journal of physiology. Cell physiology
ISSN: 1522-1563
Titre abrégé: Am J Physiol Cell Physiol
Pays: United States
ID NLM: 100901225
Informations de publication
Date de publication:
01 09 2023
01 09 2023
Historique:
pmc-release:
01
09
2024
medline:
25
8
2023
pubmed:
24
7
2023
entrez:
24
7
2023
Statut:
ppublish
Résumé
CROP-Seq combines gene silencing using CRISPR interference with single-cell RNA sequencing. Here, we applied CROP-Seq to study adipogenesis and adipocyte biology. Human preadipocyte SGBS cell line expressing KRAB-dCas9 was transduced with a sgRNA library. Following selection, individual cells were captured using microfluidics at different timepoints during adipogenesis. Bioinformatic analysis of transcriptomic data was used to determine the knockdown effects, the dysregulated pathways, and to predict cellular phenotypes. Single-cell transcriptomes recapitulated adipogenesis states. For all targets, over 400 differentially expressed genes were identified at least at one timepoint. As a validation of our approach, the knockdown of
Identifiants
pubmed: 37486064
doi: 10.1152/ajpcell.00148.2023
pmc: PMC10635647
doi:
Substances chimiques
RNA, Guide, CRISPR-Cas Systems
0
Banques de données
figshare
['10.6084/m9.figshare.22653736']
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
C648-C660Subventions
Organisme : NHLBI NIH HHS
ID : R01 HL134817
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL139478
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK125260
Pays : United States
Organisme : NIDDK NIH HHS
ID : P30 DK116074
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK106236
Pays : United States
Organisme : NHGRI NIH HHS
ID : UM1 HG011972
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL151535
Pays : United States
Organisme : NLM NIH HHS
ID : T15 LM007033
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL145708
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK120565
Pays : United States
Organisme : NIDDK NIH HHS
ID : R00 DK111916
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL156846
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK116750
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK107437
Pays : United States
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