Cas9 in vitro digestion buffer

Snippet: in EV‐based genome editing therapy. 2 MATERIALS AND METHODS 2.1 Plasmid constructs To create non‐lentiviral vectors expressing N‐myristoylated Cas9 (mCas9), Cas9/sgRNA‐empty, CRISPR vectors (OriGene, Rockville, MD, USA) were used as PCR templates. The Src(WT;8 a.a) (forward primer) and mCas9 primer (reverse primer) (Table S1) were used to obtain a PCR product, which fused the DNA sequenc

Snippet: SEPN1 KO HeLa cells were generated with the CRISPR/CAS9 technology (OriGene) and have been previously described [14]

Snippet: GFP-expressing ES2 VEGF−/− cells were generated in this study by CRISPR/Cas9 gene editing using the human VEGFA gene knockout kit (Origene). Briefly, ES2 cells were co-transfected with VEGFA sgRNA plasmid and donor vector that contains predesigned homologous sequences flanking the GFP and puromycin resistance selection cassette. Individual clones, derived from single GFP-expressing cells, were sel...

Snippet: SC71-S D5 (to stain type 2 A fiber) from Iowa Hybridoma bank. 4.12 CRISPR/CAS9 technology SELENON KD, ERO1 KD and double SELENON, ERO1KD HeLa cells were generated with the CRISPR/CAS9 technology (OriGene), closely following the manufacturer's instructions. Three different gRNAs were designed to target exon 2 of the human SELENON gene on chromosome 1. The gRNA that gave the best knock down was

Snippet: cribed by Wong, et al.39 and the Mfold web server for nucleic acid folding and hybridization prediction (http://unafold.rna.albany.edu/?q=mfold/rna-folding-form)39. Custom sgRNAs were cloned into the Origene Technologies (Rockville, MD) All-in-One CRIPSR/Cas9 vector (pCas-Guide-EF1a-GFP). For the production of the sgRNA for microinjection into embryos, a T7 in vitro transcription of the sgRNA was;...