PurposeIn vitro transcription of a mRNA encoding humanized Cas9 nuclease, with 3' UTRs suitable for germline expression in C. elegans, for doing CRISPR-Cas by RNA injection. Uses SP6 RNA polymerase.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||47911||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 3000
- Total vector size (bp) 7115
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert namehCas9 with C. elegans 5' and 3' UTRs
SpeciesH. sapiens (human), C. elegans (nematode)
- Promoter SP6
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRV (destroyed during cloning)
- 3′ cloning site KpnI (not destroyed)
- 5′ sequencing primer T3
- 3′ sequencing primer T7 (Common Sequencing Primers)
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:SP6-hCas9-Ce-mRNA was a gift from Paul Sternberg (Addgene plasmid # 47911 ; http://n2t.net/addgene:47911 ; RRID:Addgene_47911)
For your References section:Transgene-Free Genome Editing in Caenorhabditis elegans Using CRISPR-Cas. Chiu H, Schwartz HT, Antoshechkin I, Sternberg PW. Genetics. 2013 Aug 26. 10.1534/genetics.113.155879 PubMed 23979577