Misc

Protocols

• CRISPR/Cas9 Protocol [English] (pdf, 1.5MB) | Ansai and Kinoshita, 2014 (CRISPR); Ansai et al. 2014 (HMA)
  A protocol for targeted mutagenesis using CRISPR/Cas system.
• TALEN Protocol v.1.0 [in Japanese] (pdf, 1.8MB) | Ansai et al, 2013
  

sgRNA design

A design tool to select oligonucleotide sequences for pDR274 vector.

• sgRNA_design.xls (xls file, 35KB) for Microsoft Excel (except 2008 for mac)
• sgRNA_design.ods (ods file, 22KB) for Apache Open Office

Vectors

• pCS2+hSpCas9 | Ansai and Kinoshita, 2014
  [Map] (pdf, 20KB), [Sequence] (genbank, 14KB)
  This vector can be used as a template vector for Cas9 RNA using SP6 RNA polymerase.
  The hSpCas9 (human codon-optimized Streptococcus pyogenes Cas9, Cong et al, 2013) gene was cloned into the pCS2+ vector.
  RNA produced using this vector worked very well in embryos of a number of teleost fish species.
  This vector is available via Addgene (Plasmid #51815).