...coexpression EF-1 alpha AAV Deisseroth 55635 pAAV-EF1a-sCre SCre EF-1 alpha AAV Deisseroth 55636 pAAV-EF1a-Cre ... construct during the cloning process). VCre and SCre are alternative Cre recombinases that recognize ... are currently in our repository. ID Plasmid Description Promoter Expression System PI 8394 p209 pCMV-...pLOX-CW-CRE Cre CMV Lentiviral Trono 12265 pHR-CMV-nlsCRE Cre CMV Lentiviral Trono 12493 p259 pCMV-CRE-M ...coexpression Xenopus hsp70 Xenopus Ryffel 31132 pCSCre Cre CMV Xenopus Ryffel 31309 pCARCre Cre cardiac...45863 pCM157 Cre lac Bacterial Lidstrom 46388 pFosCreER Cre-ERT2;Targeting vector for Fos locus Mammalian... Mammalian Luo 47117 pT3TS-Cre for in vitro transcription of Cre lac Bacterial Ekker 47551 pDD104 (Peft...

...CRISPR plasmids for transcriptional repression of target genes....Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide like KRAB, can knock down...down gene expression by interfering with transcription. Design your gRNA to target your gene of interest...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...search the tables below for CRISPR plasmids for transcriptional inhibition. Plasmids are available for expression...

...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs... cut Goldstein 42250 DR274 T7 BsaI In vitro transcription none, need Cas9 plasmid Joung 46170 PU6::klp... plasmid Boxem 47941 pMB60 T7 BsaI In vitro transcription none, need Cas9 plasmid Boxem 47912 SP6-sgRNA-scaffold...sgRNA-scaffold SP6 AflII In vitro transcription none, need Cas9 plasmid Sternberg 177783 L4440_BioBrick-...

...in vitro transcription BsaI none S. pyogenes Mark pU6T7 71462 Mammalian/in vitro transcription BsaI none...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...Ventura pCRISPRyl 70007 Other/Yarrowia lipolytica SCR1'-tRNA yes, cut S. pyogenes LEU2 Wheeldon FgH1tUTG...Mammalian U6x2 yes, nick S. pyogenes Puro Jackson pBluescriptSKII+ U6-sgRNA(F+E) empty 74707 Mammalian hU6 none...from our collection. Plasmids Expression System Description PI Multiplex CRISPR/Cas9 Assembly System Kit ...

...efficiently block transcription (in bacteria, ~300-fold repression) by interfering with transcriptional elongation...elongation, RNA polymerase binding, or transcription factor binding. The binding specificity is determined jointly...the nuclease-deficient mutant could silence transcription on the gene expression level without genetically...targeted fluorescent genes. These plasmids are described in: Repurposing CRISPR as an RNA-Guided Platform...CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes. Gilbert LA, Larson MH, Morsut...can be ordered via the links below: ID Plasmid Description 44246 pdCas9-humanized A catalytically inactive...

... bind transcriptional co-activators and modulate transcription. The list of p53 transcriptional targets...collection of plasmids for the p53 pathway. p53 is a transcription factor and tumor suppressor activated by cellular...p53 Gene List References Background p53 is a transcription factor and tumor suppressor activated by cellular...mutations interfere with p53’s ability to activate transcription, and they also have a dominant negative effect... receptor superfamily, member 10b E2F-1 E2F transcription factor 1 Fas Fas cell surface death receptor...

...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs... Edit Cas9 H840A nickase fused to a reverse transcriptase (RT) is capable of installing targeted insertions...Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide like KRAB, can knock down...down gene expression by interfering with transcription. Design your gRNA to target your gene of interest...

...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...search the tables below for CRISPR plasmids for transcriptional activation. Plasmids are available for expression...

...Overview of tetracycline regulated inducible transcriptional activation or repression...tetracycline and not to the TRE, permitting transcription. Tetracycline-dependent promoters are developed... of VP16 (virion protein 16), an essential transcriptional activation domain from HSV (herpes simplex ...and induce your gene of interest. ID Plasmid Description Co-expressed tTA, rtTA, or TetR On or Off PI ...transactivator for your tetraycline system. ID Plasmid Description Element On or off PI 26429 pLenti CMV rtTA3 Blast...Annu Rev Microbiol . 1994;48:345-69. PubMed . Transcriptional activation by tetracyclines in mammalian cells..., tetR, rather than the tetR-mammalian cell transcription factor fusion derivatives, regulates inducible...

... the REAL-Fast assembly method (Reyon et al., manuscript submitted), and the FLASH (Fast Ligation-based...Hockemeyer et al., Nat Biotechnol. 2011 ) TALE transcriptional activators in human cells ( Maeder et al., ...identify both dimeric TALEN and monomeric TALE transcriptional activator target sites and to simplify the ...sequences of interest for potential TALEN or TALE transcriptional activator target sites and then generates individual...for FLASH assembly. Links to additional pages describing these reagents are provided below: Joung Lab ...Vectors for REAL, REAL-Fast and FLASH TALE Transcriptional Activator Expression Vectors for REAL, REAL-Fast...

.... Green Listed - a CRISPR Screen Tool Design gRNAs for custom CRISPR screens targeting a long or short...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...For more information, see the bioRxiv preprint describing ICE . Deep Sequencing Data Analysis CRISPResso... featured on Addgene's blog Protocols Lab(s) Description Plasmids in protocol Download protocol Church...and Wente Zebrafish: gRNA cloning, in vitro transcription, injection gRNA core ; Cas9 ; optimized Cas9...102.5 KB Jaenisch CRISPR plasmid summary for transcriptional activation dCas9 activators sgRNA dual expression...

...a detailed protocol description: Avidity based Extracellular Interaction Screening (AVEXIS) for the scalable...typically used in systematic protein interaction screens or cellular assays to identify novel interactions...AVEXIS (for Avidity-based extracellular interaction screen) in which the extracellular regions of receptor...are based on the pTT3 plasmid backbone which is described by Durocher et al. Nucleic Acids Research 2002.... Genome biology 2009; 10(9):R99. Large-scale screening for novel low-affinity extracellular protein interactions...

...reverse transcriptase and integrase. Once in a target cell, the virus uses the reverse transcriptase in tow...envelope and packaging plasmids. ID Plasmid Type Description PI 8449 pUMVC Packaging Contains MoMLV gag and...plasmids that have inserts. ID Plasmid 5'LTR Description PI 1780 pBABE-neo largeTcDNA MoMLV Expression...cloning and gene expression; select with puromycin or screen for GFP Bartel 32702 pMSCV-loxp-dsRed-loxp-eGFP-Puro-WPRE...see pMIG-w , a variant that has WRE, a post-transcriptional regulatory element that increases export to...

...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...replace” gene editing method in which a reverse transcriptase (RT) is fused to the C terminus of Cas9 H840A...edit is incorporated into the DNA by reverse transcription. The edited DNA strand displaces the unedited...Moloney murine leukemia virus (M-MLV) reverse transcriptase. Engineering of the PE enzyme and related components...including exploring new sources for the reverse transcriptase. Many PE enzymes and related tools are available...

...expression plasmids. Large collections or kits are described below and not included in this table. For a more...search Addgene's entire collection. ID Plasmid Description Gene/Insert PI Plasmids Expressing Worm Genes...strains. CRISPR plasmids from John Calarco's lab as described in Heritable genome editing in C. elegans via ...genome-editing events. Developed in Craig Mello's lab and described in A Co-CRISPR Strategy for Efficient Genome Editing...also been developed by the Fire laboratory and described in Efficient Marker-Free Recovery of Custom Genetic...C. elegans and other nematodes. Wormbuilder - Describes methods to engineer the genome of the genetic ...

...disease severity. The MECP2 protein is a global transcriptional regulator of thousands of genes and studies...studies have suggested roles in transcriptional repression, activity dependent de-repression, chromatin remodeling...DNA and recruit known co-repressors or other transcriptional modifiers suggests the major function of MECP2...NTD) the M ethyl B inding D omain (MBD) a T ranscriptional R epressor D omain (TRD) the TRD contains a...species (mouse or human) Mutation ID Plasmid Description Gene/Insert Mutations PI Return to top External...Developing Human Brain - includes developmental transcriptome information, prenatal LMD microarray data, ISH...

...Other Applications Screen CRISPR libraries are a powerful tool for genome-wide screening. Purify dCas9 fused...Prime Edit Cas nickase enzymes fused to reverse transcriptase install new genetic information into a specific...transposases insert large fragments of DNA. Transcriptional Regulation Activate dCas9 fused to an activator..., or fused to a repressor peptide, inhibits transcription. Epigenetics dCas9 fused to an epigenetic modifier...

...nickase system is based on the Cas9 D10A nickase described in Figure 4 of the Cong, et. al, 2013 Science ... The human U6 promoter prefers a 'G' at the transcription start site to have high expression, so adding...co-transfect in HR donor. Alternatively, a new system described recently is the double nickase system, with PX335...destroy the BbsI sites, a double digest should discriminate between positive and negative clones. Clones..., how can cells with that mutation be selected/screened? Before starting your experiment, consider co-...sequencing, for example) on each colony in order to screen for positive ones. - Or - If your HR template has...

...H134R)-EYFP Flp Yes 126080 pAAV-Ef1a-sCreDIO hChR2(H134R)-eYFP Scre No 126081 pAAV-Ef1a-vCreDIO hChR2(H134R...public folder (Link opens in a new window) of manuscripts that have used these constructs experimentally...Yes 55638 pAAV-EF1a-vCre None Yes 55635 pAAV-EF1a-sCre None No 55633 pAAV-EF1a-mCherry-IRES-Dre None No...in the Periaqueductal Gray Facilitate the Itch-Scratching Cycle via Article Tac1-Expressing Neurons in ...in the Periaqueductal Gray Facilitate the Itch-Scratching Cycle via Descending Regulation. Neuron 101(1...

...124606 SOX2-mEGFP AICSDP-60 mEGFP Transcription factor SOX-2 Transcription Factor 124607 ACTN2-mEGFP AICSDP...human cell lines of their choice. For a full description of how to use these plasmids, check out Plasmids...133963 UBTF-HaloTag AICSDP-80 HaloTag Nucleolar transcription factor UBF Nucleolus (fibrillar center) 133964...159745 TFAM-mEGFP AICSDP-72 mEGFP Mitochondrial transcription factor A (TFAM) Mitochondrial nucleoids 164499...