...knock down gene expression by interfering with transcription. Design your gRNA to target your gene of interest...at a much lower efficiency than NHEJ. ID Plasmid Gene/Insert Promoter PI Publication Browse CRISPR Plasmids...edits via homology-directed repair (HDR). ID Plasmid Gene/Insert Promoter PI Publication Prime Edit Cas9 H840A... the desired edits on an RT template. ID Plasmid Gene/Insert Promoter PI Publication Activate Catalytically...peptide can increase transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator...activator to promoter or regulatory regions of your gene of interest. If the plasmid that you choose does...dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter PI Publication Interfere Catalytically...

...shRNA expression. Gene Switch: These constructs contain two genes of interest, Genes A and B. When Cre...Cre-regulated gene expression. Depending on the construct, Cre may activate or repress gene expression. ... or “LSL” cassette) upstream of a gene of interest will prevent gene expression in the absence of Cre....the stop codon is excised, and gene expression proceeds. Cre-dependent Gene Knockout: Conversely, putting...on either side of a gene (called “floxing”, for “flanked by loxP”), will permit gene expression until Cre...Cre is absent, only Gene A is translated correctly. Cre expression excises Gene A and alters the reading...p079 pPNT6 Cre activates gene of interest, removes Neo cassette. Used in gene targeting. Mouse Targeting...

...name to find available plasmids for the gene, or browse the gene list below. For proteins with multiple...top Ras Pathway - Gene List Click on a name to find available plasmids for the gene. For components with... p53 Pathway Background Ras Pathway Plasmids Ras Gene List Resources Background Ras is a small GTPase ...a variety of pathways, which ultimately turn on genes involved in cell growth, differentiation and survival...the cell, however when mutations occur in the Ras gene it can be stuck in a constitutively active form,...multiple subunits or isoforms, individual links to each gene page are provided below. Color is used for clarity...multiple subunits or isoforms, individual links to each gene page are provided below. Symbol Name AKT AKT1 AKT2...

...knock down gene expression by interfering with transcription. Design your gRNA to target your gene of interest...at a much lower efficiency than NHEJ. ID Plasmid Gene/Insert Selectable Marker PI Publication Base Edit...>C on the opposite strand) mutations. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication...edits via homology-directed repair (HDR). ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication... the desired edits on an RT template. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication...peptide can increase transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator...activator to promoter or regulatory regions of your gene of interest. If the plasmid that you choose does...

...KRAB-MeCP2 fusion for gene and non-coding enhancer sequence repression dCas9-VPR fusion for gene activation CRISPR...CRISPR Collection The Brzezinski lab investigates gene regulation in the context of the developing mouse...based approach to knockout, repress, and activate gene targets. To accomplish this they have taken advantage...pX458 plasmid to expand its usefulness for the lab's gene regulation and developmental studies. Key plasmid...fate specification, yet differs from deleting both genes . Development. doi: 10.1242/dev.190272. PMID: 32541005...pX458 backbone ( Addgene #48138 ) dCas9 + KRAB-MeCP2 ( Addgene #110821 ) VPR ( Addgene #63798) Plasmids...Cloning Protocols Single guide cloning: Zhang Lab General Cloning Protocol (PDF, 237 KB) Dual guide cloning...

...name to find available plasmids for the gene, or browse the gene list below. For multiple isoforms or subunits...for the gene. For components with multiple isoforms or subunits, individual links to each gene page are...especially important to tumorigenesis. Inheriting only one functional copy of the p53 gene causes a large reduction...content for this page was generated with the help of . Return to top p53 Pathway - Gene List Click on a name... Ras pathway Background p53 Pathway Plasmids p53 Gene List References Background p53 is a transcription...Cancer Impact p53 is the most frequently mutated gene (>50%) in human cancer; most of these mutations ... subunits, individual links to each gene page are provided below. Color is used for clarity and does not...

... ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert Promoter...Drosophila ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Plant ID Plasmid Gene/Insert Promoter...Marker PI Publication Yeast ID Plasmid Description Gene/Insert Promoter Selectable Marker PI Publication...CRISPR nickase plasmids for the generation of single-strand breaks. CRISPR...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other...out our Suggest a Plasmid form or e-mail help@addgene.org to help us improve this resource!...

...EcTyrRS/tRNA pair (tyrS gene knocked out, tyrTV gene replaced by tolC, and tyrU gene replaced by GentR). ...EcTyrRS/tRNA pair (tyrS gene knocked out, tyrTV gene replaced by tolC, and tyrU gene replaced by GentR) and...Removed endogenous EcTyrRS/tRNA pair (tyrS gene knocked out, tyrTV gene replaced by tRNA variant that suppress...endogenous EcTyrRS/tRNA pair (tyrS gene knocked out, tyrTV and tyrU genes replaced by tRNA variant that suppress...endogenous EcTyrRS/tRNA pair (tyrS gene knocked out, tyrTV and tyrU genes replaced by tRNA variant inactivated...that contain genes with modified codons for unnatural amino acid incorporation. ID Plasmid Gene/Insert Vector...non-canonical amino acids (ncAAs). A plasmid containing the gene of interest with the modified codon (typically the...

...Return to top RNA Polymerases Addgene ID Plasmid Gene/insert 165505 pOpen-T7gene1 T7 RNA Polymerase 165516 ...165576 pOpen-T3gene1 T3 RNA Polymerase Return to top Other Enzymes Addgene ID Plasmid Gene/insert 165509... Polymerases Other Enzymes DNA Ligases Addgene ID Plasmid Gene/insert 165503 pOpen-HSlig4 DNA Ligase IV...Ligase III Return to top DNA Polymerases Addgene ID Plasmid Gene/insert 165500 pOpen-9N7polA 9 degrees N...Return to top Restriction Endonucleases Addgene ID Plasmid Gene/insert 165504 pOpen-EcoRIR EcoRI 165506...NcoI Return to top Reverse Transcriptases Addgene ID Plasmid Gene/insert 165546 pOpen-MMLV_RT (mut H) Moloney...Exonuclease III 165559 pOpen-T5gene12/13 T5 Exonuclease 165560 pOpen-T4 g32 T4 gene 32 single-stranded DNA-...

...developments with CRISPR-Cas9 gene editing have propelled both reverse and forward genetics of these parasites. ... these tools have enabled precise genetic analyses to pinpoint genes linked to parasite virulence, transmission.... Moreover, CRISPR-dCas9 gene regulation has also been established for some of these parasites. Together...at a much lower efficiency than NHEJ. ID Plasmid Gene/Insert Selectable Marker PI Publication Libraries...10 guides against 8,158 predicted protein-coding genes. Do you have suggestions for other plasmids that...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other... these efforts. The content for this page was generated with the help of Scott Lindner . Cut Fully functional...

...name to find available plasmids for the gene, or browse the gene list below. For multiple isoforms or subunits...for the gene. For components with multiple isoforms or subunits, individual links to each gene page are...this page was generated with the help of David Sabatini . Return to top mTOR Pathway - Gene List Click on...Screening Kit Background mTOR Pathway Plasmids mTOR Gene List Resources The m echanistic or m ammalian t ... subunits, individual links to each gene page are provided below. Color is used for clarity and does not...Addgene's plasmids for the mTOR pathway. enhanced signaling of mTOR, a key metabolic regulator, correlates...catabolic processes including autophagy and lysosome biogenesis. mTORC2 does not respond to nutrients, but it...

... editing multiple genes at once; using dual nickases to generate a knockout or gene edit; or using Cas9...the targeted gene. The ideal result is a loss-of-function mutation within the targeted gene. However, the...eliminate target gene function. It has also been used extensively to screen for novel genes that regulate... activate, or repress genes. Each library typically contains ∼3–6 gRNAs per gene to ensure modification...repress, visualize, and isolate genes. Activation or Repression of Target Genes Figure 9: Overview of CRISPRi...highest levels of single-gene activation (Figure 9D) In bacteria, activating gene expression is more difficult... of effective gene activators when fused with dCas9. Recently, synthetic CRISPR-Cas gene activators have...

...and aggressive plan to use gene editing, MECP2 reactivation, RNA editing, gene replacement therapy, RNA ...mutations in the MECP2 gene. Please refer to Addgene's CRISPR Guide for a general introduction to CRISPR...diagnostic criteria have disease-causing mutations in the gene methyl-CpG binding protein 2 ( MECP2 ). MECP2 Rett...caused from loss-of-function mutations in the MECP2 gene. Causative mutations for Rett syndrome typically...global transcriptional regulator of thousands of genes and studies have suggested roles in transcriptional...activity dependent de-repression, chromatin remodeling, gene activation, and more. How loss of MECP2 protein ...suggests the major function of MECP2 is to regulate gene expression either locally or globally. The MECP2...

...at a much lower efficiency than NHEJ. ID Plasmid Gene/Insert Promoter PI Publication Nick CRISPR/Cas nickase...edits via homology-directed repair (HDR). ID Plasmid Gene/Insert Promoter PI Publication Prime Edit Cas9 H840A... the desired edits on an RT template. ID Plasmid Gene/Insert Promoter PI Publication Activate Catalytically...peptide can increase transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator...activator to promoter or regulatory regions of your gene of interest. If the plasmid that you choose does...dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter PI Publication Empty gRNA Expression...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other...

... anti-CRISPR genes in bacteriophages infecting Pseudomonas aeruginosa . Anti-CRISPR genes employ varied...especially amenable to genetic screens. Pooled gRNA libraries can be used to identify genes that are important...system - utilizing various CRISPR-associated (Cas) genes to not only store a record of invading phages but...Csf1) Type VI (Cas13) Fighting Back: Anti-CRISPR Genes in Phage The CRISPR adaptive immune system seems...produces insertions/deletions disrupting a target gene. On the other hand, if a repair template with homology...according to this template, allowing for precise gene editing. These edits can include mutations seen ...given phenotype. Current libraries are available for gene knockout, as well as transcriptional activation ...

... ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert Promoter...Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Yeast ID Plasmid Gene/Insert Promoter...Selectable Marker PI Publication Zebrafish ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other...and adenine base editors (ABEs) — can be used to generate single base pair edits without double stranded...editing systems include targeted AID-mediated mutagenesis (TAM) from the Chang lab and CRISPR-X from the...out our Suggest a Plasmid form or e-mail help@addgene.org to help us improve this resource!...

...for the EGFP Reporter Gene Guide RNA expression plasmids for endogenous human genes Guide RNA expression...these vectors for efficiently modifying endogenous genes in zebrafish (Hwang and Fu et al., Nat Biotechnol...expression plasmids for endogenous zebrafish genes...

...RNA-mediated gene silencing mechanism. It is a useful tool to investigate the roles of specific genes or to ...to knockdown genes with potentially harmful mutations in therapeutic settings. As a tool in mammalian cell...Lentiviral vectors are a preferred method for long-term gene silencing. Once the viral vector is taken up by ...RNA (shRNA): design, delivery, and assessment of gene knockdown . Methods in Molecular Biology, 629 , ...Additional Resources Addgene Resources pLKO.1 cloning vector protocol Other Addgene viral protocols Lentiviral...mammalian RNA interference (RNAi) experiments at Addgene; browse by constitutive lentiviral shRNA, cre-regulated...

...listed below. When targeting other gene loci, you will prepare the gene-specific CRISPR and donor vectors...multiple genes. Plasmids can be found associated with the following article: Paix, et al. Genetics 2014 Allen...CRISPR plasmids for endogenous tagging of your gene of interest. CRISPR...repair template design. How to use CRISPR to tag your gene of interest Mendenhall and Myers Tagging System ... the plasmids as listed in each row. If your own gene of interest is currently unavailable, you will need...terminal affinity tag (3xFLAG-2xSTREP) on endogenous genes for the isolation of native protein complexes. This... is different from the conventional HDR-mediated gene knock-in. The detailed procedure can be found in...

...Plasmids Find Plasmids by Gene Name Alternative Gene Names Gene Description AR plasmids AR, RP11-383C12.1...and results in activation or repression of target gene expression. Nuclear receptors play a key role in...homeostasis, and the receptors and their downstream genes are associated with numerous disease pathways. In...this collection, email deposit@addgene.org to be paired with an Addgene scientist. Nuclear Receptor Plasmids...collaboration with research labs from around the globe, Addgene has assembled a Nuclear Receptor plasmid collection...