Other

Dataset Information

0

Base editing mutagenesis maps functional alleles to tune human T cell activity


ABSTRACT: CRISPR-enabled genetic screening is a powerful tool to discover genes that control T cell function and has nominated candidate target genes for immunotherapies1–6. However, new approaches are required to probe specific nucleotide sequences within key genes. Systematic mutagenesis in primary human T cells could discover alleles that tune specific phenotypes. DNA base editors are powerful tools to introduce targeted mutations with high efficiency7,8. Here, we develop a large-scale base editing mutagenesis platform with the goal of pinpointing nucleotides encoding amino acid residues that tune primary human T cell activation responses. We generated a library of ~117,000 sgRNAs targeting base editors to protein coding sites across 385 genes implicated in T cell function and systematically identified protein domains and specific amino acid residues that regulate T cell activation and cytokine production. We discovered a broad spectrum of alleles with variants encoding critical residues (in PIK3CD, VAV1, LCP2, PLCG1 and DGKZ and others), comprising both gain-of-function and loss-of-function mutations. We validated the functional effects of diverse alleles and further demonstrated that base edit hits could positively and negatively tune T cell cytotoxic function. Finally, higher-resolution screening using a base editor with relaxed PAM requirements9 (NG versus NGG) revealed specific structural domains and protein-protein interaction sites that can be targeted to tune T cell functions. Base editing screens in primary immune cells provide biochemical insights with potential to accelerate immunotherapy design.

ORGANISM(S): Homo sapiens

PROVIDER: GSE244774 | GEO | 2023/10/11

REPOSITORIES: GEO

Dataset's files

Source:
Action DRS
Other
Items per page:
1 - 1 of 1

Similar Datasets

| PRJNA1024532 | ENA
2024-01-12 | GSE253184 | GEO
2024-01-12 | GSE253182 | GEO
2021-10-18 | GSE181109 | GEO
2019-09-25 | GSE136749 | GEO
2024-01-23 | GSE253655 | GEO
2022-08-24 | GSE204833 | GEO
2024-09-17 | GSE262442 | GEO
| PRJNA918720 | ENA
2021-07-21 | GSE180351 | GEO