The immediate 4-bp sequence upstream of the transcriptional start site was changed to ACTT in all U6 constructs to simplify the cloning procedures.Ĭloning vector for Staphylococcus aureus Cas9 (SaCas9) guide RNA transcription, controlled by the U6 snRNA promoter #3 (3’ of Cre08.g371751). The 20 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. reinhardtii antibiotic resistance gene insertion (aphVII, aphVIII, ble).įrom Andre Greiner, Peter Hegemann lab, Humboldt University-Berlin October 2017Ĭloning vector for Staphylococcus aureus Cas9 (SaCas9) guide RNA transcription, controlled by the U6 snRNA promoter #2. HindIII and KpnI restriction sites can be used for C. The immediate 4-bp sequence upstream of the transcriptional start site was changed to ACTT in all U6 constructs to simplify the cloning procedures. 8:2281-2308Ĭloning vector for Staphylococcus aureus Cas9 (SaCas9) guide RNA transcription, controlled by the U6 snRNA promoter #1 (5’ of Cre08.g377251). Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F (2013) Genome engineering using the CRISPR-Cas9 system. Targeting of photoreceptor genes via zinc-finger nucleases and CRISPR/Cas9 in Chlamydomonas reinhardtii. Visit for more info or contact A., Kelterborn, S., Evers, H., Kreimer, G., Sizova, I., and Hegemann, P. Overview of all CRISPR/Cas9 plasmids from the Hegemann lab From Simon Kelterborn, Peter Hegemann lab, Humboldt University-Berlin October 2017Ĭloning vector for Streptococcus pyogenes Cas9 (SpCas9) single guide RNA transcription, controlled by the U6 snRNA promoter #4 (3’ of Cre15.g640800).
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