CRISPR/Cas 9 Plasmids for use in Xenopus
Addgene is working with the leading scientists in the field to assemble the reagents and information you need to use the CRISPR/Cas9 technology in your own lab. The following Cas9 and gRNA expression plasmids have been designed for use in Xenopus.
Fully functional Cas9 enzymes designed to introduce a double-strand break (DSBs) at a specific location based on a co-expressed gRNA-defined target sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ); a mechanism which frequently causes insertions or deletions (InDels) in the DNA, possibly resulting in frameshifts. If a repair template with high homology to the DNA surrounding the DSB is introduced along with the Cas9 and gRNA plasmids, the cell may instead repair the break using homology-directed repair (HDR). HDR is much less error-prone than NHEJ and can be used to faithfully introduce specific genomic changes.
|Plasmid||Gene/Insert||Promoter||Selectable Marker||PI||Publication||Hidden Extra Search Info|
|pCS2-3xFLAG-NLS-SpCas9-NLS||3xFLAG-NLS-SpCas9-NLS||CMV||Chen||Efficient RNA/Cas9-mediated genome editing in Xenopus tropicalis. Development. 2014 Jan 8.||Cas9 expression plasmid pCS2+|
Empty gRNA Expression Vectors
Select a gRNA expression plasmid based on factors such as selectable marker or cloning method. When using the CRISPR/Cas system, you will need to express both a Cas9 protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas9 act as an all-in-one vector, but their function (cut, nick, activate, interfere...) is limited to that of the Cas9 present on the plasmid. gRNA plasmids that do not co-express Cas9 require a separate plasmid that does so; however, these independent gRNA plasmids can be paired with a wide variety of Cas9 plasmids and therefore are not limited to a single Cas9 function.
|Delivery||Resistance||Co-expressed Cas9||Depositing lab|
|pUC57-Simple-gRNA backbone||T7||BsaI||In vitro transcription||none, need Cas9 plasmid||Chen|
Last Updated: February 20, 2015