.... When a repair template is present, the cell may repair a DSB using homology-directed repair (HDR) instead...be used with a repair template to introduce specific edits via homology-directed repair (HDR). ID Plasmid...gRNA-defined target sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ...genomic changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the...Zebrafish Xenopus CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Base...base editors convert adenine to inosine, which is replaced by guanosine to create A->G (or T->C on the opposite...nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous end joining (...

.... When a repair template is present, the cell may repair a DSB using homology-directed repair (HDR) instead...be used with a repair template to introduce specific edits via homology-directed repair (HDR). ID Plasmid...Zebrafish Xenopus CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The...gRNA-defined target sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ...genomic changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the...base editors convert adenine to inosine, which is replaced by guanosine to create A->G (or T->C on the opposite...nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous end joining (...

...generate TALORs (TAL Orthongal Repressors) that can be used to custom repress gene expression in yeast. TALORs... customizable arrays of polymorphic amino acid repeats in the TAL effectors. View Addgene's TALEN Guide...efficiently assemble TALEN constructs with custom repeat arrays. TALEN Kits TALEN Kit Description Depositing...along with strong nuclear localisation tags and repress transcription initiation when targeted to DNA sequences... pFUS array vectors, destination vectors and a reporter vector for mammalian cell-based validation assay...N- and C-terminal domains (N153AA, C47AA). The reporter vector pGL4-SSA can be used for single-strand ... TALEs), 63 wild-type TAL amino acids after the repeat domain, and C-terminal SV40 NLS and HA tags. Plasmid...

..., Addgene is distributing ready-to-use viral preparations in engineered systemic serotypes from the Viviana...and full sequencing of the final viral vector preparation. Systemic Delivery AAV Capsid Choice Need help...Addgene’s Data Hub and the Caltech CLOVER Center’s repository of published capsid use (Link opens in a new ...: Systemic Capsids PHP.eB These viral vector preparations were produced with the pUCmini-iCAP-PHP.eB plasmid...tropism exhibited by the PHP.eB serotype has been reported to occur by using the cellular receptor LY6A (...expression iCre AIBS , Ting PHP.S These viral vector preparations were produced with the pUCmini-iCAP-PHP.S plasmid...tdTomato Control Boyden PHP.V1 These viral vector preparations were produced with the pUCmini-iCAP-PHP.V1 plasmid...

...tetracycline-dependent repression. Mutating these residues led to the development of a reverse Tet repressor, or rTetR...derived from pCW57.1. rtTA was replaced with tTA, and Puromycin was replaced with Blasticidin selection. ...to be replaced by the gene of interest. tTA Off Sabatini 92099 AAVS1_Puro_Tet3G_3xFLAG_Twin_Strep Bidirectional...regulated inducible transcriptional activation or repression Plasmid... the Tet Response Element (TRE) A TRE is seven repeats of a 19 nucleotide tetracycline operator (tetO)...sequence, and is recognized by the tetracycline repressor (tetR). In the endogenous bacterial system, if...presence of tetracycline for induction, rather than repression. The new transactivator rtTA ( r everse t etracycline-controlled...

.... When a repair template is present, the cell may repair a DSB using homology-directed repair (HDR) instead...be used with a repair template to introduce specific edits via homology-directed repair (HDR). ID Plasmid...gRNA-defined target sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ...genomic changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the...Zebrafish Xenopus CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Base...base editors convert adenine to inosine, which is replaced by guanosine to create A->G (or T->C on the opposite...nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous end joining (...

.... When a repair template is present, the cell may repair a DSB using homology-directed repair (HDR) instead...be used with a repair template to introduce specific edits via homology-directed repair (HDR). ID Plasmid...Zebrafish Xenopus CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The...gRNA-defined target sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ...genomic changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the...base editors convert adenine to inosine, which is replaced by guanosine to create A->G (or T->C on the opposite...nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous end joining (...

...example: To activate or repress a target gene using dCas9-activators or dCas9-repressors, gRNAs should be targeted...region where the edit should be made HDR requires a repair template and displays reduced efficiency compared...Base editors can make a limited set of mutations. Repress or Interfere (CRISPRi) Reduce expression of a particular...without permanently modifying the genome dCas9-repressor (such as dCas9-KRAB) or dCas9 gRNA(s) targeting...promoter is typically used for gRNA May contain reporter gene (e.g. GFP) to identify and enrich positive...vector or separate transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify...CRISPR protocols from Addgene depositors JOVE Video Reprint: Protocol for genomic deletions in mammalian cell...

...and high-titer viral preparations of plasmids that are currently in our repository. ID Plasmid Description...sites AAV Uchida Cre Reporters and Tools In this subtype of loxP plasmids, reporter genes indicate which...Mammalian Pelczar 62732 Cre Reporter DsRED and EGFP Cre recombinase reporter Lentiviral Geijsen 65726 pLV-CMV-LoxP-DsRed-LoxP-eGFP...Vectors Cre-containing Plasmids loxP Constructs Cre Reporters & Tools Additional Resources References Background...Fluorescent Cre: The fusion of Cre to a fluorescent reporter enables visualization of Cre expression. Optimized... of choice. We also offer ready-to-use viral preparations of select Cre expression plasmids. Addgene's...HRE:GFP-pGK:Cre Cre and hypoxia-responsive GFP reporter HRE Lentiviral Jacks 69570 pAAV-EF1a-N-CretrcintG...

...Plasmid Description PI Viral Preps Addgene distributes ready-to-use viral preps made from plasmids in our... BRAIN Initiative collection of plasmids, viral preps, and antibodies here. Collections...the research community. Browse plasmids , viral preps , and antibodies created with support from the BRAIN...use. Learn more about our viral service ID Viral Prep Description PI 65417-AAV8 pAAV-hSyn-dF-HA-KORD-IRES-mCitrine...plasmid encoding for Archon1 fluorescent voltage reporter, Cre-dependent expression Edward Boyden 108912...Expresses the genetically-encoded fluorescent norepinephrine(NE) sensor GRAB_NE1m in neurons Yulong Li 123308...Expresses the genetically-encoded fluorescent norepinephrine(NE) sensor GRAB_NE1m in neurons Yulong Li 123309...

...Envelope and Packaging Plasmids Reporter Plasmids A few examples of reporter plasmids that can be used for... Lentiviral reporter vector expressing firefly luciferase. pCMV-FLuc - Retroviral reporter vector expressing...vesicular stomatitis virus G) are packaged as replication-defective virus particles with the SARS-CoV-2...available in standard format and as a Lentiviral prep . ID Plasmid Description Mutations Industry PI Due...pHAGE-CMV-Luc2-IRES-ZsGreen-W - Lentiviral dual reporter vector expressing firefly luciferase and ZsGreen...ZsGreen. NL4-3 mCherry Luciferase - Lentiviral dual reporter vector expressing mCherry and firefly luciferase...destabilized EGFP. Lenti-luciferase-P2A-Neo - Lentiviral reporter vector expressing firefly luciferase with neomycin...

.... When a repair template is present, the cell may repair a DSB using homology-directed repair (HDR) instead...be used with a repair template to introduce specific edits via homology-directed repair (HDR). ID Plasmid...Zebrafish Xenopus CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The...gRNA-defined target sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ...genomic changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the...nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous end joining (...

...amino acid repeats. These repeats only differ from each other by two amino acids, their repeat-variable ... of 18 TAL effector repeats. The kit can be used to make arrays from 12-31 repeats. Perhaps one of the... to this DNA targeting ideal. Figure 1: (A) Representation of a zinc finger (ZF) nuclease. Zinc fingers...if not slightly lower. Figure 2: Simplified representation of the Voytas/Bogdanove Golden Gate TALEN kit... would expect that assembling relatively small, repeat-laden DNA regions into a single construct would...from Addgene , custom arrays consisting of 12-31 repeats can be assembled and inserted into a variety of...

...- and serine-containing peptide linker (GGSGGS) repeat pET28CLY2 Peptide linker standard consisting of...of ECFP and EYFP connected by 2 GGSGGS repeats pET28CLY3 Peptide linker standard consisting of ECFP and...and EYFP connected by 3 GGSGGS repeats pET28CLY4 Peptide linker standard consisting of ECFP and EYFP connected... connected by 4 GGSGGS repeats pET28CLY5 Peptide linker standard consisting of ECFP and EYFP connected...connected by 5 GGSGGS repeats pET28CLY6 Peptide linker standard consisting of ECFP and EYFP connected by 6 GGSGGS...GGSGGS repeats pET28CLY7 Peptide linker standard consisting of ECFP and EYFP connected by 7 GGSGGS repeats...consisting of ECFP and EYFP connected by 8 GGSGGS repeats pET28CLY9 Peptide linker standard consisting of...

...tyrS gene knocked out, tyrTV gene replaced by tolC, and tyrU gene replaced by GentR). Expresses archaeal ...tyrTV gene replaced by tRNA variant that suppress the TAG nonsense codon, and tyrU gene replaced by tRNA ...tyrS gene knocked out, tyrTV gene replaced by tolC, and tyrU gene replaced by GentR) and release factor 1...biopolymers. Tips for Success Before beginning to reprogram the genetic code, there are several things to ... are true. Always express first with a control reporter gene – GFP for E. coli or mCherry-GFP for mammalian...RF1 function removed, with Ubiquitin-UAG-sfGFP reporter George Church 98565 C321.ΔClpS.Ub-UAG-sfGFP all...removed, ClpS inactivated, with Ubiquitin-UAG-sfGFP reporter George Church 174513 Syn61 No TCG, TCA, or TAG...

...Selectable markers Viral expression and packaging Reporters, shRNA expression, transgenics and genome...host organism. Host Relevant Promoters Representative Empty Backbones Mammalian CMV, SV40, EF1a, ...plasmid series - Expression with Strep, His or optimized YFP tags for purification...Tag or Fusion Protein Common uses Representative Empty Backbones Flag Epitope tag... Marker Typical Host Organism Representative Empty Backbones Neomycin (G418) ...methods: Delivery method Advantages Representative Empty Backbones Lentiviral Can transduce... AdEasy®-1 plasmid Back to Top Reporters, shRNA expression, transgenics and genome ...

...modifications: replacement of the Cbh promoter with EF1alpha addition of an mCherry reporter variant nuclear...utilizes a CRISPR/Cas9 based approach to knockout, repress, and activate gene targets. To accomplish this ...advantage of the Zhang lab all-in-one (guide/Cas9/reporter) pX458 plasmid family. They have made several ...nuclear reporters via an H2B fusion shuttle plasmid for shuttling U6-guide cassettes to make dual guide ...fusion for gene and non-coding enhancer sequence repression dCas9-VPR fusion for gene activation CRISPR/Cas9...175573 and 175574 express dCas9-KRAB-MeCP2 for repression. Plasmids 175571 and 175572 express dCas9-VPR...

...2~3-fold repress on targeted fluorescent genes. These plasmids are described in: Repurposing CRISPR as...Design Tools CRISPR Blog Posts We have recently repurposed the bacterial immune system, CRISPR ( C lustered...pathway, as an RNA-guided DNA binding platform, to repress expression of arbitrary genes in bacteria or human...efficiently block transcription (in bacteria, ~300-fold repression) by interfering with transcriptional elongation...plasmids in bacteria could cause up to 300-fold repress on targeted genes. 2. Two-plasmid CRISPRi system...Lentiviral vector that contains sgRNA targeting repetitive sequence of human MUC4 exon 3 46915 pU6-sgGAL4...

.... When a repair template is present, the cell may repair a DSB using homology-directed repair (HDR) instead...Zebrafish Xenopus CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Fully...gRNA-defined target sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ...genomic changes, researchers use ssDNA or dsDNA repair templates with 1. homology to the DNA flanking ...

.... When a repair template is present, the cell may repair a DSB using homology-directed repair (HDR) instead...Zebrafish Xenopus CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The...gRNA-defined target sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ...genomic changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the...