pX330-U6-Chimeric_BB-CBh-hSpCas9

Protocol tips

Upstream tips	Protocol tips	Downstream tips
To access a plasmid, keep the plate on dry ice to prevent thawing. Using a sterile pipette tip (20uL or 200uL one), puncture the seal above an individual well and spread a portion of the glycerol stock onto an agar plate.

Upstream tips
To access a plasmid, keep the plate on dry ice to prevent thawing. Using a sterile pipette tip (20uL or 200uL one), puncture the seal above an individual well and spread a portion of the glycerol stock onto an agar plate.

Publication protocol

CRISPR/Cas9 for PrPC knockout
The human PrPC gene sequence (NM_000311.3) was used to design the guide RNA using the Optimized CRISPR Design (http://crispr.mit.edu/). gRNA sequences selected were: Hu PrPC (Top1) CACCGgctgggggcagccgatacccg/Hu PrPC (Bottom1) AAACcgggtatcggctgcccccagcC. The gRNAs were phosphorylated, annealed, and cloned into px330-U6-GFP vector (Addgene) according to the Addgene website instructions. The construct was sequenced, transfected into the U251 cell line with lipofectamine 2000 (Invitrogen) following the manufacturer’s instructions, and the clones were isolated by serial dilution.

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Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing CRISPR Human - Repression PrPC using pX330-U6-Chimeric_BB-CBh-hSpCas9 from Addgene.

Manufacturer protocol

Download the product protocol from Addgene for pX330-U6-Chimeric_BB-CBh-hSpCas9 below.

Download manufacturer protocol

Videos

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