Protocols for use are provided in the publications associated with the collection cited below, and the supplementary material of these publications. This includes a detailed description on how to generate CRISPR/Cas vectors containing zCas9i by MoClo (1st set of plasmids, Grützner et al: Figures S14-17) and a laboratory manual for using pDGE plasmids (2nd set of plasmids, Barthel et al: Appendix 1). See the PDF below for an overview of the plasmids included in this kit.
How to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
“The zCas9i cloning kit: MoClo-compatible CRISPR/Cas9 cloning kit with intronized Cas9 was a gift from Sylvestre Marillonnet and Johannes Stuttmann (Addgene kit #1000000171).”
For your Reference section:
High-efficiency genome editing in plants mediated by a Cas9 gene containing multiple introns Grützner R, Martin P, Horn C, Mortensen S, Cram EJ, Lee-Parsons CWT, Stuttmann J, Marillonnet S. Plant Communications 2021 Volume 2, Issue 2 doi: 10.1016/j.xplc.2020.100135. Publication.
Highly efficient multiplex editing: One-shot generation of 8x Nicotiana benthamiana and 12x Arabidopsis mutants Stuttmann J, Barthel K, Martin P, Ordon J, Erickson JL, Herr R, Ferik F, Kretschmer C, Berner T, Keilwagen J, Marillonnet S, Bonas U. Plant J. 2021 Feb 12. doi: 10.1111/tpj.15197. Epub ahead of print. PubMed PMID: 33577114.