...Resistance Activity PI 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP... dCAS-VP64_Blast dCAS9 (D10A, N863A) Blasticidin 3rd generation lenti vector encoding dCAS9-VP64 with ...are made from plasmids that are currently in our repository. You can obtain the plasmid from us or receive... list of gRNA plasmids targeting specific genes. For more information on CRISPR plasmids and function,...Accessories for Activating Gene Expression Catalytically-dead Cas9 (dCas9) can be fused to a transactivator...with 2A Blast resistance marker (EF1a-NLS-dCas9(N863)-VP64-2A-Blast-WPRE) Zhang 61426 lenti MS2-P65-HSF1_... of available viruses based on plasmids in our repository that express fluorescent proteins. ID Name Insert...

...BFP fusion expression plasmid 44248 pgRNA-humanized A customizable gRNA expression plasmid for use with...46912 pMSCV-LTR-dCas9-VP64-BFP Human expression vector containing MSCV LTR promoter, dCas9 that is fused...to 2x NLS, VP64 and tagBFP 51023 pSLQ1658-dCas9-EGFP Human expression vector containing dCas9 that is fused...customizable gRNA expression plasmid for use with the bacterial Cas9 constructs 46910 pHR-SFFV-dCas9-BFP Human...pHR-SFFV-dCas9-BFP-KRAB Human expression vector containing SFFV promoter, dCas9 that is fused to 2x NLS, tagBFP...46913 pMSCV-LTR-dCas9-p65AD-BFP Human expression vector containing MSCV LTR promoter, dCas9 that is fused...1. Two-plasmid CRISPRi system for bacterial gene knockdown The first plasmid ( pdCas9_bacteria ) contains...

...2. pmax dCas9 Activator Expression Plasmids ID Plasmid 48223 pAC91-pmax-dCas9VP64 : dCas9VP64 on pmax ... Plasmid 48236 pAC2-dual-dCas9VP48-sgExpression : Dual expression construct expressing both dCas9VP48 ... the sgRNA expressing fragment produced by PCR), or expressed from a single dual-expression vector [48236...Individual plasmids can be ordered via the links below: Table 1. dCas9 Activators sgRNA Dual Expression ID Plasmid...puro-selectable) on pmax expression vecor. Table 3. dCas9 Activators Gateway Donors ID Plasmid 48214 pAC1-pCR8-... pAC152-dual-dCas9VP64-sgExpression : Dual expression construct expressing both dCas9VP64 and sgRNA from... pAC153-dual-dCas9VP96-sgExpression : Dual expression construct expressing both dCas9VP64 and sgRNA from...

...tracrRNA: This plasmid contains three expression cassettes. To target a given site, the plasmid can be digested...Addgene plasmids #61422 - #61424 were designed for transient transfection : 61422 : Expresses dCAS9-VP64 ...nuclei in addition to sgRNA expression. This plasmid contains two expression cassettes, Cre recombinase... SpCas9 Plasmids The CRISPR/Cas system can be implemented in mammalian cells by co-expressing the S. pyogenes...., Science 2013). Additionally, we have a plasmid expressing SpCas9 alone without sgRNA. Full references...nickase) + single guide RNA: These plasmids contain two expression cassettes, a human codon-optimized ...

...repressor (such as dCas9-KRAB) or dCas9 gRNA(s) targeting promoter elements of target gene dCas9-KRAB is more ...permanently modifying the genome dCas9-activator (such as dCas9-VP64) gRNA(s) targeting promoter elements...CRISPRi) Reduce expression of a particular gene(s) without permanently modifying the genome dCas9-repressor ...effective than dCas9 alone for mammalian cell lines. Activate (CRISPRa) Increase expression of an endogenous...activators exist, including the multi-plasmid SAM system. Select Expression System To use CRISPR, you will need...to the transient Cas9 expression. The table below summarizes the major expression systems and variables...Delivery method Expression System Components of System Application Mammalian expression vector Cas enzyme...

...Mammalian Lentiviral expression of Cas9, dCas9, or dCas9-VP64 along with 1-4 sgRNAs expressed from independent... on the plasmid. gRNA plasmids that do not co-express a Cas protein require a separate plasmid that does...Vectors CRISPR Plasmids: Empty gRNA Expression Vectors Browse CRISPR Plasmids By Function Genome Editing Cut...maintained by the Doudna Lab . Plasmid Expression System Cloning Enzyme Co-expressed Cas9 Cas9 System Selection...can be used to express multiple gRNAs from a single plasmid with or without expression of Cas9. Highlighted.... Can express Cas9 with up to 3 gRNAs. Frew pSQT1313 Mammalian Single plasmid for the expression of multipe...requires expression of Csy4 (eg Plasmid 53369 ). Joung pX333 Mammalian Single all-in-one plasmid for tandem...

...system (Figure 9C) SAM activators - co-expression of dCas9-VP64 with a modified scaffold gRNA and additional...tools require constitutive expression of the dCas9 fusion and/or expression of multiple fusions. The effects...achieved by using orthogonal dCas9s (e.g. S. pyogenes dCas9 and S. aureus dCas9) tagged with different fluorescent...biotin tagging of dCas9 by fusing a biotin acceptor site to dCas9 and co-expressing BirA biotin ligase...multiple gRNAs using a single plasmid ensures that all gRNAs will be expressed in the same cell. This is ...genomic DNA. dCas9 has also been used for genome-wide screens to activate or repress gene expression in mice...human cells. The simplest dCas9-based activators and repressors consist of dCas9 fused directly to a single...