Variant-EFFECTS Pipeline

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Jesse Engreitz

Stanford Center for Connecting DNA Variants to Function and Phenotype

Analysis of IL2RA promoter 5-bp tiling mutagenesis edit effects on IL2RA expression that were measured by Variant-EFFECTS (formerly known as Variant-FlowFISH) in Jurkat cells stimulated for 24h with PMA and anti-CD3 antibody.

Jesse Engreitz, Stanford

Homo sapiens Jurkat cell line, stimulated with 100 ng/mL phorbol 13-acetate 12-myristate for 24 hours, 5 μg/mL T-cell surface glycoprotein CD3 gamma chain for 24 hours, modified with CRISPR prime editing Sp-nCas9-M-MLV RT (PE2), transfected with a guide library, sorted on expression of IL2RA

Homo sapiens (male) Jurkat cell line treated with 100 ng/mL phorbol 13-acetate 12-myristate for 24 hours, 5 μg/mL T-cell surface glycoprotein CD3 gamma chain for 24 hours, modified with CRISPR prime editing Sp-nCas9-M-MLV RT (PE2)

Treatment of 100 ng/mL phorbol 13-acetate 12-myristate for 24 hours

Treatment of 5 μg/mL T-cell surface glycoprotein CD3 gamma chain for 24 hours

Homo sapiens (male) Jurkat cell line (sorting details: FACS bin for 51-75% expression of IL2RA) treated with 100 ng/mL phorbol 13-acetate 12-myristate for 24 hours, 5 μg/mL T-cell surface glycoprotein CD3 gamma chain for 24 hours, modified with CRISPR prime editing Sp-nCas9-M-MLV RT (PE2), transduced (lentivirus) with a guide library

Homo sapiens (male) Jurkat cell line (sorting details: FACS bin for 0-25% expression of IL2RA) treated with 100 ng/mL phorbol 13-acetate 12-myristate for 24 hours, 5 μg/mL T-cell surface glycoprotein CD3 gamma chain for 24 hours, modified with CRISPR prime editing Sp-nCas9-M-MLV RT (PE2), transduced (lentivirus) with a guide library

Homo sapiens (male) Jurkat cell line (sorting details: FACS bin for 76-100% expression of IL2RA) treated with 100 ng/mL phorbol 13-acetate 12-myristate for 24 hours, 5 μg/mL T-cell surface glycoprotein CD3 gamma chain for 24 hours, modified with CRISPR prime editing Sp-nCas9-M-MLV RT (PE2), transduced (lentivirus) with a guide library

Homo sapiens (male) Jurkat cell line (sorting details: FACS bin for 26-50% expression of IL2RA) treated with 100 ng/mL phorbol 13-acetate 12-myristate for 24 hours, 5 μg/mL T-cell surface glycoprotein CD3 gamma chain for 24 hours, modified with CRISPR prime editing Sp-nCas9-M-MLV RT (PE2), transduced (lentivirus) with a guide library

CRISPR prime editing Variant-EFFECTS targeting IL2RA: variant effects

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Audit Category: Internal Action

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