...integration of donor DNA. Genome-Wide Screens Using CRISPR In a genetic screen, a population of cells with mutations...Targeting dCas9 to transcription start sites is sufficient to repress transcription by blocking initiation..., G. M. (2013). CAS9 transcriptional activators for target specificity screening and paired nickases for...in your lab, from knockouts to pooled library screening. Science...CRISPR Basics Small Edits Large Edits Genome-Wide Screening Cas9 Fusion Tools Cas9 Alternatives Resources ...approaches like zinc finger nucleases (ZFNs) or transcription-activator-like effector nucleases (TALENs) required...technologies, making CRISPR perfect for genome-wide screens. This guide will provide a basic understanding ...

...The collection can be used for screening transcription factors for cellular programming or cancer drug ...simplifies identification of the transcription factor isoform in pooled screens. This collection and library...library can be used for screening transcription factors for applications such as cellular programming or cancer...for NGS QC and bulk screening (PDF, 85 KB) TF ORF barcode dial out PCR for 10X scRNA-seq (PDF, 59.2 KB)...to simplify identification during screening. Library ID PI Description MORF library 137000, 192821, 1000000218...Browse the Zhang lab’s comprehensive human transcription factor collection, available at Addgene. The...Library The MORF library is a comprehensive human transcription factor collection that contains 1,836 genes ...

...Access to the Paper & Protocol Video Download the manuscript here: Bauer et al, 2014 322.2 KB If you have ...production of genomic deletions. In addition, we describe techniques for deletion, identification, and characterization...referenced Tables and Figures, please access the manuscript PDF posted at the top of the page. CRISPR Design... the genomic recognition site. NOTE: Figure 1 describes possible deletion strategies for genes and non-coding...leading to nonsense mediated decay of the mRNA transcript ( Figure 1B ). Use the example guides for the...be 24-mer oligos ( Table 4 ). Design Deletion Screening Primers Design one set of primers internal to ...deletion. For this approach, the internal primers described in step 2.1 may be omitted. CRISPR Cloning Anneal...

... luciferase. Transcriptional Element Screening Optimized STARR-seq ( S elf- T ranscribing A ctive R egulatory...promoter/enhancer regions. Blue/white screening can be used for screening of transformants in cloning process...including transcriptional reporters for NF-kb, TGF-b, c-Myc, p53 and MAPK/JNK transcriptional reporters...collection of luciferase plasmids for assays of transcriptional regulation and bioluminescent reporters. Plasmid... protein half-lives, making them useful as transcriptional reporters. However, they are not secreted, ...plasmids : Luminescence-based protein-protein screening method for the systematic and quantitative analysis...egulatory R egion) plasmids : Genome-wide screening of enhancer activity using a Firefly luciferase reporter...

...below. Return to top Cas9 SAM transcriptional activation plasmids and screening library CRISPR/Cas9 Synergistic...RNA 48137 : PX165; SpCas9 Return to top GeCKO Screening Library GeCKO are pooled lentiviral CRISPR libraries...and mouse) for performing genome-wide knockout screens. The libraries are available in two formats: 1 ...) is an engineered protein complex for the transcriptional activation of endogenous genes. It consists...database (23,430 isoforms) for gain-of-function screening. For more information, please visit the Addgene... references are below. Available plasmids are described below: 61591 : PX601; CMV-driven SaCas9; U6-driven...sgRNA to a constitutively Cas9-expressing mouse. Described here are AAV vectors that can be combined with...

...: dCas9 fused to transcriptional activators or repressors can modulate transcription in a reversible manner...palindromic repeat sequences. (1) The CRISPR array is transcribed to make the pre-CRISPR RNA (pre-crRNA). (2) The...in vitro. In 2013, Cong et al. and Mali et al. described CRISPR-based genome editing in mammalian cell ... of new applications. The first CRISPR papers described two main categories of genome edits. First, the... locus makes it especially amenable to genetic screens. Pooled gRNA libraries can be used to identify ...are available for gene knockout, as well as transcriptional activation or repression. Combined with the...sequencing, CRISPR is a robust system for genome-wide screening. CRISPR Technological Advances CRISPR technology...

...mutant cells that are then screened for a phenotype of interest. Screening experiments using a pooled ...CRISPR pooled libraries for large-scale functional screens. Available for knockout, activation, inhibition...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...Blog Posts CRISPR is a useful tool for genetic screening experiments, due to the relative ease of designing... gRNAs for each target gene. In a CRISPR screening experiment, target cells are treated with the ...libraries and keep the following in mind: CRISPR screening experiments require electroporation to amplify...

... Plasmid Description Industry mEmeralds Plasmid Description Industry Reds Plasmid Description Industry... Plasmid Description Industry Cyans Plasmid Description Industry Oranges Plasmid Description Industry ...Industry Greens Plasmid Description Industry Yellows Plasmid Description Industry Photoactivatables Plasmid Description... Description Industry Do you have suggestions for other plasmids that should be added to this list? Fill...

...pSerOTS-C1* (V70) 188537 ). These reagents are described in detail in (Mohler et al., Mol Syst Biol 2023...at the central position. Introducing Hi-P for screening phosphorylation-dependent protein-protein interactions...NEDD4-2; 111705-8 ) is available from Addgene. Screening kinases. The mode #1 phosphosite library can also...Phosphorylated Isomers (iSPI). This new resource described in (Gassaway et al., Nature Methods 2022) is adapted...Iterative Synthetically Phosphorylated Isomers (iSPI) described in: A multi-purpose, regenerable, proteome-scale...in a new window) rEcoli XpS and a new pSerOTS described in: System-wide optimization of an orthogonal ..., 551 KB) Human Serine Phosphopeptide Library described in: Encoding human serine phosphopeptides in bacteria...

... ID Plasmid Depositing PI Description Golden Gate TALEN 2.0 38142 RCIscript-GoldyTALEN Daniel Carlson ...new tools for genome engineering. Fusions of transcription activator-like (TAL) effectors of plant pathogenic...with custom repeat arrays. TALEN Kits TALEN Kit Description Depositing PI FusX TALEN Assembly System Contains...expression of TALENs from a truncated CAGs promoter. RCIscript-GoldyTALEN is designed for in vitro synthesis ...strong nuclear localisation tags and repress transcription initiation when targeted to DNA sequences within...destination vectors can be used to create TALE-transcription factors and are used at the final assembly step...TALEN kit, (ii) a lacZ fragment for blue/white-screening in E.coli , (iii) CAG promoter and Kozak sequence...

...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...-dU6:3gRNA dU6:3 BbsI Injection or in vitro transcription Virmilion none, need Cas9 plasmid Bullock and... dU6:1 and dU6:3 BbsI Injection or in vitro transcription Virmilion none, need Cas9 plasmid Bullock and...-dU6:1gRNA dU6:1 BbsI Injection or in vitro transcription Virmilion none, need Cas9 plasmid Bullock and...

...at the bottom of this page. For more in depth descriptions of each serotype, check out Addgene's blog on...Browse Available PHP.eB AAV ID Name Promoter Description Category PI Controls 28306 pAAV-FLEX-tdTomato... Browse Available PHP.S AAV ID Name Promoter Description Category PI 28306 pAAV-FLEX-tdTomato CAG tdTomato...Browse Available PHP.V1 AAV ID Name Promoter Description Category PI 104052 pAAV-CAG-DIO-EYFP CAG EYFP...Browse Available MaCPNS1 AAV ID Name Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control...Browse Available MaCPNS2 AAV ID Name Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control...Browse Available CAP-B10 AAV ID Name Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control...

...testing transcription factor binding to reporter plasmid pH3U3-zif268 (omega) . Transcription factors ... answer many interesting questions. Does my transcription factor bind to a particular sequence? Do these...for easy western blotting or staining, and RNA transcript tags to monitor RNA localization. Using these...variety of inducible promoters, their associated transcription factors, and reporter genes. pCS-PesaRlux 47640...Lactose/IPTG inducible expression of EsaR, a transcription factor responsive to 3OC6HSL. EsaR derivatives...that element. When this activation requires a transcription factor to bind to a small molecule, the reporter...derivatives as a fusion to RNA polymerase subunit ω. Transcription factor binding to a putative promoter region...

...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...via homology-directed repair (HDR). ID Plasmid Description Gene/Insert Promoter Selectable Marker PI Publication...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...direct dCas9 to a specific locus, avoiding known transcription factor and other protein binding sites. ID Plasmid...

... Plasmids expressing Cas12a/Cas13a, Reverse Transcriptases, etc. that may be useful for detection assays... industry is continually growing. ID Plasmid Description Industry PI Return to top Other Coronavirus Plasmids... SARS-CoV, MERS, BAT-CoV Plasmids ID Plasmid Description Industry PI Due to export control regulations...plasmids related to these genes below. ID Plasmid Description Industry PI For more information on these genes...2020.06.23.166397 (Link opens in a new window) reverse transcriptase, protease, and Bst polymerase constructs now...unpublished) Taq polymerase and MMLV reverse transcriptase expression plasmids from the Drew Endy and Philippa...2020.03.29.013342 (Link opens in a new window) Reverse Transcriptase plasmids from Andrew Ellington's lab. High-surety...

...find more information. Delivery Method Species Description Article PI Lentivirus Human Expression of human...human Klf4, Oct4, c-Myc, and Sox2 as VP-16 transcriptional activating fusions in single lentiviral vectors...human iPSC generation OCT4 and SOX2 Work as Transcriptional Activators in Reprogramming Human Fibroblasts...Progenitor Cells Lentiviral Human Cooperative Transcription Factor Induction Mediates Hemogenic Reprogramming...Oligodendrocyte Progenitor Cells Lentiviral Mouse Transcription factor-mediated reprogramming of fibroblasts...fibroblasts into functional astrocytes by defined transcription factors. Stem Cell Reports. 2015 Jan 13;4(1)...Neurons Lentiviral Human Chemical modulation of transcriptionally enriched signaling pathways to optimize the...

...expression in plants: Maize Transcription Factor ORFeome - John Gray. Rice Transcription Factor ORFeome - John... Addgene Blog: Plant Biology CRISPR Cloning Transcription Factors RNAi Empty Backbones Fluorescent Proteins...plant transformation vectors - Daniel Voytas. Transcription Factors The following collections can be used...John Gray. Plant transcription factors for Agrobacterium-mediated transient expression or plant transformation...promoter or cloning method of choice. ID Plasmid Description Promoter Cloning Method PI Return to top Fluorescent...or fluorescent protein of choice. ID Plasmid Description Promoter PI Return to top Other Resources Addgene...Genomics Resource (Link opens in a new window) - Transcriptomic and metabolomic resources for 14 key medicinal...

...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...direct dCas9 to a specific locus, avoiding known transcription factor and other protein binding sites. ID Plasmid...

...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...direct dCas9 to a specific locus, avoiding known transcription factor and other protein binding sites. ID Plasmid...

...Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases Other Enzymes (Link opens in...polymerases, RNA polymerases, DNA ligases, reverse transcriptases, and restriction enzymes. These enzymes are... cloning, isothermal amplification, reverse transcription of RNA into DNA, and dNTP synthesis. Additionally...Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases Other Enzymes DNA Ligases Addgene...165568 pOpen-NcoIR NcoI Return to top Reverse Transcriptases Addgene ID Plasmid Gene/insert 165546 pOpen-MMLV_RT...Moloney Murine Leukemia Virus (MMLV) Reverse Transcriptase (RNAse H deactivated by 3 mutations) 165556 ...Moloney Murine Leukemia Virus (MMLV) Reverse Transcriptase RNaseH - (lacking RNaseH domain) 165566 pOpen-TthRT...