... plasmids dCas9-KRAB-MeCP2 fusion for gene and non-coding enhancer sequence repression dCas9-VPR fusion...175574 express dCas9-KRAB-MeCP2 for repression. Plasmids 175571 and 175572 express dCas9-VPR for activation... Plasmid and Sequence Derivative Sources Original pX458 backbone ( Addgene #48138 ) dCas9 + KRAB-MeCP2...activation. Usage Guide sequences can be designed using Benchling or using similar design tools and ordered...Golden Gate cloning to insert the desired guide RNA sequence by digestion of the backbone with BbsI followed...phosphorylated oligonucleotides containing the guide target sequence: Up to two guides can be expressed from the same...

... protein dCas9 is expressed along a CRISPR array that contains 20 bp "addresses", where dCas9 is going...going to bind on the chromosome. When dCas9 binds to a gene, it blocks the progression of the RNA polymerase...Mismatches between the guide RNA and the target sequence lower the repression level. By controlling the...for quick integration of the aTc-inducible pTet-dCas9 in the attachment site of the phage 186. Detailed... Bikard, D., & Van Teeffelen, S. (2018). Tuning dCas9’s ability to block transcription enables robust,...Schematic of the strain expressing two reporters and dCas9 under a P-tet promoter integrated in the chromosome...be controlled using a plasmid‐borne CRISPR array coding for guide RNAs (diamonds) interspaced with CRISPR...

...of a specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory ...PI Publication Interfere Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide... promoter/enhancer or the beginning of the coding sequence. If the plasmid you’re using does not also ...immunoprecipitate FLAG-tagged Cas9. Design your gRNA sequence to direct dCas9 to a specific locus, avoiding known transcription...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...specific location based on a gRNA-defined target sequence. DSBs are preferentially repaired in the cell ...Marker PI Publication Base Edit Catalytically dead dCas9 fused to a cytidine deaminase protein becomes a ...

...target have high sequence identity to the spacer sequence and contain a PAM sequence just downstream of...of a specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory ...PI Publication Interfere Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide... promoter/enhancer or the beginning of the coding sequence. If the plasmid you’re using does not also ...immunoprecipitate FLAG-tagged Cas9. Design your gRNA sequence to direct dCas9 to a specific locus, avoiding known transcription...Promoter PI Publication 64325 3xFLAG-dCas9/p-bacteria 3xFLAG-dCas9 pLtetO-1 Fujii Efficient isolation of...specific location based on a gRNA-defined target sequence. DSBs are preferentially repaired in the cell ...

...Design your gRNA sequence to direct the dCas9-FP fusion to a specific genomic sequence - potential target...of a specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory ...PI Publication Interfere Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide... promoter/enhancer or the beginning of the coding sequence. If the plasmid you’re using does not also ...both enCHIP and CAPTURE, design your gRNA sequence to direct dCas9 to a specific locus, avoiding known transcription...specific location based on a gRNA-defined target sequence. DSBs are preferentially repaired in the cell ...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...

...Tools CRISPR Blog Posts Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide... promoter/enhancer or the beginning of the coding sequence. If the plasmid you’re using does not also ...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors... separate gRNA expression plasmid to target the dCas9-repressor to your specific locus. Want more information...

...evaluation of their effect on the coding sequence, functional noncoding elements and off-target sites. This...also accepted) from a deep sequencing experiment a reference amplicon sequence to assess and quantify the...input to assess HDR frequency. An sgRNA sequence (without PAM sequence) can be provided, to compare the predicted...to the position of the observed mutations. Coding sequence/s may be provided to quantify frameshift and...thermophilus Cas9, or Cpf from your input sequence or list of sequences. RGEN (RNA-guided endonuclease) Tools...Cas-OFFinder identifies gRNA target sequences from an input sequence and checks for off-target binding....Japan; Identify candidate gRNA target sequences in an input sequence, which can be an accession number, ...

...Description 47948 3xFLAG-dCas9/pCMV-7.1 enChIP plasmid, expresses 3xFLAG-dCas9 in mammalian cells for enChIP...in mammalian cells 51240 3xFLAG-dCas9/pMXs-puro Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis...regions of interest 51258 3xFLAG-dCas9/pMXs-IG Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis...regions of interest 51259 3xFLAG-dCas9/pMXs-I2 Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis...regions of interest 51260 3xFLAG-dCas9/pMXs-neo Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis...human IRF-1 promoter 62190 3xFLAG-dCas9/pTEF1p-CYC1t Expresses 3xFLAG-dCas9 in budding yeast for enChIP analysis...neomycin resistance gene 64325 3xFLAG-dCas9/p-bacteria Expresses 3xFLAG-dCas9 in E. coli in a tetracycline-dependent...

...of a specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory ...PI Publication Interfere Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide... promoter/enhancer or the beginning of the coding sequence. If the plasmid you’re using does not also ...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...specific location based on a gRNA-defined target sequence. DSBs are preferentially repaired in the cell ...Marker PI Publication Base Edit Catalytically dead dCas9 fused to a cytidine deaminase protein becomes a ...Marker PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can...

...permanently modifying the genome dCas9-repressor (such as dCas9-KRAB) or dCas9 gRNA(s) targeting promoter elements... elements of target gene dCas9-KRAB is more effective than dCas9 alone for mammalian cell lines. Activate...without permanently modifying the genome dCas9-activator (such as dCas9-VP64) gRNA(s) targeting promoter elements...select a target sequence and design your gRNA. Know your cell line/organism and genomic sequence When possible...should sequence the region you are planning to modify prior to designing your gRNA, as sequence variation...activate or repress a target gene using dCas9-activators or dCas9-repressors, gRNAs should be targeted to...identifying all PAM sequences within the genetic region to be targeted. If there are no PAM sequences for your chosen...

...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...forward genetics of these parasites. Moreover, CRISPR-dCas9 gene regulation has also been established for some...specific location based on a gRNA-defined target sequence. DSBs are preferentially repaired in the cell ... with 10 guides against 8,158 predicted protein-coding genes. Do you have suggestions for other plasmids...

...target site sequence (20bp) and need to be followed on the 3' end by a 3bp NGG PAM sequence. We have found...target site sequence (30bp) and need to be followed on the 3' end by a 3bp NGG PAM sequence. 3. SpCas9 ...target site sequence (21-22bp) and need to be followed on the 3' end by NGRRT or NGRRN PAM sequences. We have... vector encoding dCAS9-VP64 with 2A Blast resistance marker 61426 : lentiviral vector encoding the MS2...of CRISPR-associated (Cas) genes as well as non-coding RNA elements capable of programming the specificity...human genome-wide library to activate all known coding isoforms form the RefSeq database (23,430 isoforms... for transient transfection : 61422 : Expresses dCAS9-VP64 activator with 2A GFP 61423 : Expresses the...

...achieved by using orthogonal dCas9s (e.g. S. pyogenes dCas9 and S. aureus dCas9) tagged with different fluorescent... be any ~20 nucleotide sequence, provided it meets two conditions: The sequence is unique compared to ...(PAM) The PAM sequence (NGG) serves as a binding signal for Cas9, but the exact sequence depends on which... target, the seed sequence (8–10 bases at the 3′ end of the gRNA targeting sequence) will begin to anneal...matters. Mismatches between the target sequence in the 3′ seed sequence inhibit target cleavage, while mismatches...determined by both the gRNA sequence and the Cas9 enzyme. Ideally, a gRNA targeting sequence will have perfect ... genetic edits. The dCas9 molecule binds to target DNA based on the gRNA sequence without cleavage. This...

...Resistance Activity PI 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A ...dCAS-VP64_Blast dCAS9 (D10A, N863A) Blasticidin 3rd generation lenti vector encoding dCAS9-VP64 with 2A ...resistance marker (EF1a-NLS-dCas9(N863)-VP64-2A-Blast-WPRE) Zhang Pooled Barcoding Libraries ID Name Description...56,762 unique sgRNAs. This backbone does not contain dCas9. 56,762 unique sgRNAs targeting 18,885 human genes...non-targeting controls. This backbone does not contain dCas9. 57,050 unique sgRNAs targeting 18,901 human genes...non-targeting controls. This backbone does not contain dCas9. 57,011 unique sgRNAs targeting 18,899 human genes...by an RNA guide (gRNA) that harbors targeting sequences. Browse our repository for a list of gRNA plasmids...

... enabling CRISPR/dCas9-interference (CRISPRi) modulation of enhancers and non-coding RNAs in zebrafish... for plasmids that contain a zebrafish gene or sequence . The table below highlights plasmids that contain...Zebrafish Genome Project - Access the latest reference sequence assembly for the zebrafish genome. Zebrafish Model...zebrafish. CRISPRz - A database of CRISPR/Cas9 target sequences that have been experimentally validated in zebrafish...Periklis Pantazis Lab Multibow: digital spectral barcodes for cell tracing - Sean Megason Lab Zebrabow -...tagging - Michel Bagnat lab. Targeting cassettes encoding fluorescent proteins for tagging genes at endogenous... Gavin Wright Lab. Collections of plasmids that encode zebrafish receptors or monoclonal antibodies to...

...of a specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory ...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...specific location based on a gRNA-defined target sequence. DSBs are preferentially repaired in the cell ...Promoter PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can... separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene...target site by homology to a genomic DNA locus and encodes a primer binding site and the desired edits on ...

...Regulation Activate dCas9 fused to an activator peptide activates gene expression. Interfere dCas9 alone, or fused...protocols, find software for gRNA design and deep sequencing analysis, discover links to CRISPR forums, and...repressor peptide, inhibits transcription. Epigenetics dCas9 fused to an epigenetic modifier alters chromatin...powerful tool for genome-wide screening. Purify dCas9 fused to an epitope tag can be used to isolate a...libraries for knockout, activation, inhibition, and barcoding experiments. Browse CRISPR Plasmids by Function...

...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...pooled library and the use of Next-Generation Sequencing (NGS) to confirm library representation. CRISPR... Zhang N/A 15 Varies CRiNCL - Human CRISPRi Non-coding Libraries 86538 — 86550 Inhibition Human Weissman...Human Paired-guide RNA (pgRNA) Library for Long Non-coding RNAs (lncRNAs) 89640 Knockout Human Wei 3rd Varies...Human Mali 3rd 1 74 Perturb-seq Guide Barcodes (GBC) 85968 Barcode Human Weissman 3rd N/A N/A Retroviral...Available for knockout, activation, inhibition, and barcoding. CRISPR... Addgene's collection. Library Type Activation Barcode Knockout Inhibition Base Editing Prime Editing ...

...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors... pegRNA directs the Cas9 nickase to the target sequence where it nicks the non-target strand and generates..., if the edited 3’ flap is excised, the target sequence remains unchanged and available as a substrate...nickase to the target site. The longer pegRNA also encodes a primer binding site (PBS) and the desired edits...

...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...gRNA(s) to guide the Cas9 protein to your target sequence, as well as design and clone in the homology arms...2A-PuroR cassette, flanked by microhomologous sequences and gRNA target sites. The first deposited PITCh...produces a single plasmid targeting vector that encodes both a guide RNA transcript and a repair template...

...Does my transcription factor bind to a particular sequence? Do these two proteins interact? What happens ...localization: Contains a 2x array of the binding sequence for RNA-binding protein, MS2. Can be used in combination...localization: Contains a 2x array of the binding sequence for RNA-binding protein MS2 fused to dsRED. Can...Tac (lactose/IPTG inducible) MBP, 6x His, signal sequence for localization to the periplasm David Waugh ...N-terminal 6x His tag and an MBP tag. Contains a signal sequence to localize the protein of interest to the periplasm...of N and C-terminal epitope tags, localization sequences, and cleavage sites that can be used to express...Assembly Accessory Reagents Kit Various Plasmids Sequence chosen by researcher Beta-galactosidase Keith ...