Purpose / Description: Cloning vector for Streptococcus pyogenes Cas9 (SpCas9) single guide RNA transcription, controlled by the U6 snRNA promoter #4 (3’ of Cre15.g640800)
Reference: Targeting of Photoreceptor Genes in Chlamydomonas reinhardtii via Zinc-Finger Nucleases and CRISPR/Cas9
Promoter: U6 snRNA promoter #4 (3’ of Cre15.g640800)
Resource information:
Cloning vector for *Streptococcus pyogenes* Cas9 (SpCas9) single guide RNA transcription, controlled by the U6 snRNA promoter #4 (3’ of Cre15.g640800). The 2 bp target site can be inserted in a cut-ligation reaction as annealed oligos into an Esp3I cloning site following the protocol of Ann Ran (Ran et al. 213). The immediate 4-bp sequence upstream of the transcriptional start site was changed to ACTT in all U6 constructs to simplify the cloning procedures.