id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-003945-esnyjoq5	Hu, Zheng	Customized one-step preparation of sgRNA transcription templates via overlapping PCR Using short primers and its application in vitro and in vivo gene editing	2019-10-24		.txt	text/plain	2317	166	58	title: Customized one-step preparation of sgRNA transcription templates via overlapping PCR Using short primers and its application in vitro and in vivo gene editing However, conventional methods of generating sgRNA either direct synthesis sgRNA or requires plasmid construction for PCR amplification to make transcription templates [7] , which is either expensive or cumbersome and requires several steps to produces gRNAs. Cell & Bioscience In the present study, we described a simplified and customized in vitro synthesis approach utilizing multiple overlapping primers to synthesize DNA fragment for six selected gene loci as the transcription templates of sgR-NAs by a single-step sequential primer extension under appropriate conditions, and tested the cleave efficiency to targeted loci by using these sgRNAs combined with Cas9 nuclease in vitro and in vivo. Summary, we demonstrated an approach by combining CRISPR/Cas9 gene editing technology with multiple overlapping primers to synthesize DNA fragment as the template of sgRNA to substitute the conventional method.	./cache/cord-003945-esnyjoq5.txt	./txt/cord-003945-esnyjoq5.txt