PurposeExpress LdPBK_241510.1 and LdMT targeting gRNAs simutaneously
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||84292||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Total vector size (bp) 3900
Vector typeCRISPR ; Leishmania donovani
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameLdBPK_241510.1 targeting gRNA and LdMT targeting gRNA
gRNA/shRNA sequenceGGTGTCGTTCTGCAGCCA and GTCCAGGCGCAGTGGATGC
GenBank IDLdBPK_241510.1 in TritrypDB LdBPK_131590.1 in TritrypDB
- Promoter L. donovani ribosome RNA promoter
- Cloning method Restriction Enzyme
- 5′ cloning site Bbs I (destroyed during cloning)
- 3′ cloning site Bbs I (destroyed during cloning)
- 5′ sequencing primer SP6 Promoter
- 3′ sequencing primer T7 promoter (Common Sequencing Primers)
Terms and Licenses
This dual gRNA expression plasmid is used to target both LdBPK_241510.1 and LdMT gene. In the article, it was used to generate the targeted chromosome translocations between chromosome 13 and 24 and to enrich the edited LdBPK_241510.1 mutants by selection with miltefosine. For addgene customers, this dual gRNA expression plasmid will mainly be used as the PCR template to generate new double gRNAs expression vector or co-targeting MT vector. See article for more detail.
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:pSPneogRNA241510+MT was a gift from Greg Matlashewski (Addgene plasmid # 84292 ; http://n2t.net/addgene:84292 ; RRID:Addgene_84292)
For your References section:Optimized CRISPR-Cas9 Genome Editing for Leishmania and Its Use To Target a Multigene Family, Induce Chromosomal Translocation, and Study DNA Break Repair Mechanisms. Zhang WW, Lypaczewski P, Matlashewski G. mSphere. 2017 Jan 18;2(1). pii: e00340-16. doi: 10.1128/mSphere.00340-16. eCollection 2017 Jan-Feb. mSphere00340-16 [pii] PubMed 28124028