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 Primary Airway Epithelia (Human)
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Results

Gene_editing_in_vitro_by_various_peptide_variants_delivering_Cas_RNPs_to_primary_Human_airway_epithelia_cells.__
Editing Efficiency
Record Id Condition Editor Model Delivery Target Locus Guide Dosage Editing Efficiency % indel Image
15000000535 S10, Cas12a, CFTR AsCas12a (Feldan Therapeutics) Primary Airway Epithelia (Human) S10 CFTR g-45_C12a 1.33uM Cas12a/2.0uM gRNA/40uM Shuttle Peptide
15000000536 S18, Cas12a, CFTR AsCas12a (Feldan Therapeutics) Primary Airway Epithelia (Human) S18 CFTR g-45_C12a 1.33uM Cas12a/2.0uM gRNA/40uM Shuttle Peptide
15000000537 S85, Cas12a, CFTR AsCas12a (Feldan Therapeutics) Primary Airway Epithelia (Human) S85 CFTR g-45_C12a 1.33uM Cas12a/2.0uM gRNA/40uM Shuttle Peptide
15000000538 CM18-PTD4, Cas12a, CFTR AsCas12a (Feldan Therapeutics) Primary Airway Epithelia (Human) CM18-PTD4 CFTR g-45_C12a 1.33uM Cas12a/2.0uM gRNA/40uM Shuttle Peptide
15000000539 S10, Cas12a, HPRT AsCas12a (Feldan Therapeutics) Primary Airway Epithelia (Human) S10 HPRT g-38330_C12a [RNP]: 2.5 µM; [gRNA]: 2.0 μM; [Peptide]: 40uM
15000000540 S18, Cas12a, HPRT AsCas12a (Feldan Therapeutics) Primary Airway Epithelia (Human) S18 HPRT g-38330_C12a [RNP]: 2.5 µM; [gRNA]: 2.0 μM; [Peptide]: 40uM
15000000541 S85, Cas12a, HPRT AsCas12a (Feldan Therapeutics) Primary Airway Epithelia (Human) S85 HPRT g-38330_C12a [RNP]: 2.5 µM; [gRNA]: 2.0 μM; [Peptide]: 40uM
15000000542 CM18-PTD4, HPRT AsCas12a (Feldan Therapeutics) Primary Airway Epithelia (Human) CM18-PTD4 HPRT g-38330_C12a [RNP]: 2.5 µM; [gRNA]: 2.0 μM; [Peptide]: 40uM
15000000543 S10, Cas9, CFTR SpCas9 (Feldan Therapeutics) Primary Airway Epithelia (Human) S10 CFTR g-11_C9 [RNP]: 2.5 µM; [gRNA]: 2.0 μM; [Peptide]: 40uM
15000000544 S18, Cas9, CFTR SpCas9 (Feldan Therapeutics) Primary Airway Epithelia (Human) S18 CFTR g-11_C9 [RNP]: 2.5 µM; [gRNA]: 2.0 μM; [Peptide]: 40uM
15000000545 S85, Cas9, CFTR SpCas9 (Feldan Therapeutics) Primary Airway Epithelia (Human) S85 CFTR g-11_C9 [RNP]: 2.5 µM; [gRNA]: 2.0 μM; [Peptide]: 40uM
15000000546 CM18-PTD4, Cas9, CFTR SpCas9 (Feldan Therapeutics) Primary Airway Epithelia (Human) CM18-PTD4 CFTR g-11_C9 [RNP]: 2.5 µM; [gRNA]: 2.0 μM; [Peptide]: 40uM

Publication Title
Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia. NCBI

Shuttle peptides deliver Cas12a and Cas9 RNPs to HAE

Shuttle peptides deliver Cas12a and Cas9 RNPs to HAE. A) Schematic showing CFTR locus in region of 3849 + 10C>T mutation (not to scale) and the sequence of the Cas12a guide RNA target. B) Editing at the CFTR locus following delivery of Cas12a RNPs using four different peptides. Shuttle peptides were tested for Cas12a RNP delivery using gRNA targeting CFTR intron 22–23. Materials were applied for 15 min, cells were harvested 72 hr later for Surveyor assay; Indel% determined by Sanger sequencing. Asterisks denote bands observed with gene editing. Np indicates Cas12a RNP with no peptide. c) S10 peptide dose–response on Cas12a RNP editing of CFTR locus. HAE transduced with fixed RNP concentration [Cas12a]: 1.33 µM; [gRNA]: 2 µM and S10 peptide concentrations varied (20–50 μM). Cells incubated with peptide-RNP for 15 min, and harvested 72 h later for Surveyor assay (Control: Cas12a RNP alone). D) Effect of incubation time and repeated of peptide-Cas12a RNP delivery on editing. [S10]: 40 μM; [RNP]: 40 µM, applied for indicated times. After 72 h, cells prepped for Surveyor assay and Sanger sequencing (Np indicates Cas12a RNP with no peptide, incubated for 3 h; Rpt denotes repeated application of peptide/RNP × 3 daily doses). n = 3 donors. E) Schematic of HPRT1 locus and Cas12a guide RNA target sequence along with editing efficiency on delivery of RNPs. Screen of four peptide formulations at 40 μM concentration, [RNP]: 2.5 µM; [gRNA]: 2.0 μM on primary HAE. Indicated peptide-RNP applied for 3 h; 72 hr later, cells were processed for Surveyor assay. Asterisks denotes genome editing. n = 3 donors. F) Schematic of the CFTR locus and Cas9 guide target (exon 11) and editing efficiency in HAE after Cas9 RNP delivery with each of four shuttle peptides. The same four peptide formulations were applied at [40 μM], with [RNP]: 2.5 μM; [gRNA]: 2.0 μM. Indicated shuttle peptide and Cas9 RNP applied for 3 h; 72 h later, cells processed for Surveyor assay. Asterisks denote genome editing. n = 3 donors. Pubmed