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Experiment: Testing newly chemically modified crRNA and tracrRNA to activate the TLR reporter in human cells

PI:   Erik J Sontheimer, PhD 
 
 

Chemically modified crRNA and tracrRNA were delivered by electroporation in presence of amphiphilic peptide to transgenic human HEK-293T cells harboring the TLR-MCV1 reporter. Gene editing was determined by reporter activation.
Editing Assay: - Calculated editing efficiency by flow cytometry analysis of RFP expression.

Filter Results ..
Editors
 SpyCas9-3xNLS
Guide Target Locus
 TLR MCV reporter
  Guides
 STS118 (TLR-MCV1a; Sp_t2:Sp_c20_TLR_MCV1a)
 STS119 (TLR-MCV1b; Sp_t2:Sp_c20_TLR_MCV1b)
Models
 HEK-293T-disrupted_GFP with MCV-mcherry-Puro
Sex
 Female

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Results

Testing_newly_chemically_modified_crRNA_and_tracrRNA_to_activate_the_TLR_reporter_in_human_cells
Editing Efficiency
Record Id Condition Sex Editor Model Delivery Target Locus Guide Dosage Editing Efficiency % of RFP+ cells Image
15000000547 STS118 (TLR-MCV1a) (5 pmol RNP) Female SpyCas9-3xNLS HEK-293T-disrupted_GFP with MCV-mcherry-Puro FSD117D1 TLR MCV reporter STS118 (TLR-MCV1a; Sp_t2:Sp_c20_TLR_MCV1a) 5 pmol RNP
15000000548 STS118 (TLR-MCV1a) (20 pmol RNP) Female SpyCas9-3xNLS HEK-293T-disrupted_GFP with MCV-mcherry-Puro FSD117D1 TLR MCV reporter STS118 (TLR-MCV1a; Sp_t2:Sp_c20_TLR_MCV1a) 20 pmol RNP
15000000549 STS119 (TLR-MCV1b) (20 pmol RNP) Female SpyCas9-3xNLS HEK-293T-disrupted_GFP with MCV-mcherry-Puro TLR MCV reporter STS119 (TLR-MCV1b; Sp_t2:Sp_c20_TLR_MCV1b) 20 pmol RNP