We narrowed to 102 results for: cas9 expression construct

...cloning and expressing your gene of interest with GFP expression.  Cloning and expressing RNAi pMKO.1 ...Plasmid #44246) Expression of a catalytically inactive (dead), human codon-optimized Cas9 under the control...find more lentiviral expression vectors (empty backbones) here. Cloning and expressing new shRNA sequences...lentiviral backbone from the Zhang lab expressing S. pyogenes Cas9 and a gRNA cassette. It can be used to...Lentiviral Expression System Kit (Kit #1000000060) The MuLE (Multiple Lentiviral Expression) system facilitates...Cre-dependent gene expression, gene knockout, and selection marker removal. CRISPR technology pdCas9-humanized ...inducible gene overexpression, knockdown (shRNA, miR-30-shRNA), mutation/editing (CRISPR-Cas9) or deletion...

...death of transduced cells and loss of transgene expression (Interestingly, the strong immunogenicity of ...research and its therapeutic potential. Note: constructs discussed are based on Adenovirus type 5. Three...capacity is limited to 8.2 kb, and in vivo transgene expression ceases relatively quickly due to immune response... capacity (up to 36 kb!), long-term transgene expression and negligible toxicity. However, their production... cells can be broken down into 5 key steps: Construction of the rAdV plasmid (~1 week): The AdEasyTM ...popular method for creating adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector ...generate the final recombinant adenoviral plasmid construct, please see Addgene’s Adenoviral Guide webpage...

...cells express iCre with a mitochondrial anchor and a TEV cleavage site. Recipient cells express DsRed ...Affordable, low-tech CRISPR/Cas9 experiments By Ashley Waldron CRISPR/Cas9 has revolutionized biotechnology...chromoprotein and gRNA plasmids. Nuclease-dead Cas9 (dCas9) protein is supplied separately. Results can ...cell-surface proteomes (Wu et al., 2025). The team expressed extracellularly-anchored horseradish peroxidase...nuclease, and excises the DsRed-STOP to trigger expression of EGFP. Figure 2: Schematic of the MitoTRACER...Confocal image of CA2 neurons in a mouse brain slice expressing PinkyCaMP; scale bar 50 µm. Reproduced from Fink...and hindbrain regions of live zebrafish larvae expressing FRCaMPi or SomaFRCaMPi in neurons; scale bars...

...resources for a full selection of plasmids expressing Cas9 and empty gRNA backbones. A few examples of... Using CRISPR/Cas9 - Jeffrey Essner Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) expression plasmids - Keith...Lab CRISPR/Cas9-based conditional mutagenesis in zebrafish - Wenbiao Chen Lab A CRISPR/Cas9 Vector System...fusion expression in organisms or cells in which an exogenous mRNA can be injected and expressed. EXPRESs...in zebrafish. Zebrafish expression atlas - A zebrafish-specific gene expression database that provides ...modified CRISPR-Cas9 system - Shuo Lin Lab Re-purposed Ac/Ds transgenic system enabling CRISPR/dCas9-interference...morpholinos, zinc finger nucleases, TALENS, and CRISPR/Cas9-based gene editing make the zebrafish a stalwart...

...pluripotent stem cell (iPS) technology and CRISPR-Cas9 constructs from Addgene to gene-edit mutations into XP...looking at a bunch of different strategies with CRISPR-Cas9. There are so many different variants and strategies...ATP1A3. The gene encodes a sodium-potassium pump expressed specifically in neurons of the brain. McSweeney... of those mutations, they now plan to use CRISPR-Cas9 technology to create cell lines and/or mouse models...ve proposed to use plasmids with modified nickase Cas9 protein in order to target the specific gene region...

...the ssDNA construct as described in Figure 2.  You can also use Addgene plasmids to purify Cas9 protein.... 47327 pET-28b-Cas9-His 6xHis (C terminal); NLS (C terminal) 62933 pET-Cas9-NLS-6xHis 6xHis... Find Plasmids for Cas9 Purification Find CRISPR Plasmids for Mammalian Expression ...mice pre-CRISPR was very arduous. First, you’d construct a plasmid with your desired changes, long homology.... recommend ribonucleoprotein (RNP) complexes of Cas9 protein and the separated guide RNA components (...increase T7 promoter activity. To avoid re-cutting by Cas9 after editing, make sure that the repair template... terminal); NLS (C terminal) 62934 pET-NLS-Cas9-6xHis 6xHis (C terminal); NLS (N terminal) ...

...proteins, including Cas9 and dCas9-transcription factors, for which low expression levels are optimal....these two systems can be coexpressed for sophisticated control of gene expression. An E. coli light-sensitive... by Tabor’s lab, making light-controlled gene expression in E. coli easier and more robust. Synthetic...engineer new pathways with sophisticated gene expression controls. Most cell types do not respond to low-intensity... common E. coli histidine kinase. This hybrid construct senses light, and an obligate chromophore allows...only about 10-fold, precluding the study of gene expression at very high or very low levels. Schmidl et al... to better fine-tune the level of output gene expression based on the input light intensity. In addition...

...protein can then be co-expressed in essentially any transgenic zebrafish line expressing a GFP-tagged version...display increased neuronal expression in the brain while having decreased expression in the liver after systemic...only AAV.CAP-B10 causes brain wide expression, with no liver expression, after retro-orbital sinus vein ...Increased neuronal expression using Cre-mediated AAV Targeted Evolution method (M-CREATE) dCas9-SSAP with reduced...10.1038/s41593-021-00969-4.   Increased neuronal expression using Cre-mediated AAV Targeted Evolution method...vein injection of a CAG-driven mNeonGreen construct in mouse. Image from Goertsen D, et al., 2022.  Find...of up to 20%, comparable to commonly used CRISPR/Cas9 methods for large inserts. However, this system ...

...to edit a gene, the expression of Cas9 is no longer necessary. However, Cas9 can be toxic if continuously...leading to transient expression of gene products (Wu et al., 2004). Over time, the expression diminishes as ...2008). 2. Transient expression of transgenes/shRNAs If you want your cells to express a protein or nucleic...the expression of the delivered payload is gradually reduced. For example, after using CRISPR/Cas9 to ...continuously expressed (Fu et al., 2013). Using IDLVs ensures transient expression without the risks associated...associated with prolonged expression. 3. Stable expression in non-dividing cells IDLVs can also be useful for...like creating stable cell lines, delivering CRISPR/Cas9 to cells for genome editing, and introducing inducible...

...lab, if you read a paper and saw an interesting construct you wished to use, you could either make it yourself...Addgene makes it so much easier to share your constructs from lab to lab. Promega supports the Addgene...endogenous tagged genes generated through CRISPR/Cas9 knock-in. Finally, NLuc is very inviting for endogenous...only 19 kDa. An example is the CRISPaint-NLuc construct (Plasmid #67178) for use in the system outlined...fusions LumiFluor Reporters. The LumiFluor reporter constructs were designed to be packaged into a retrovirus...reporter was used to sort successfully infected/expressing cells by flow cytometry. The isolated cells were...reporter can improve deep tissue imaging. The constructs for GpNLuc and OgNLuc are available through Addgene...

...Blainey’s lab has developed a multiplexing approach to Cas9-based pooled CRISPR screens called CROPseq-multi...using selection by Puromycin, Zeocin®, or nuclear expression of BFP2.     Figure 1: CROPseq-multi ...produced 90% accurate integration of multiplexed constructs into target genomes and a 10-fold improvement...Anzalone et al., 2019, including modifications to the Cas9 or reverse transcriptase enzymes, inhibition of ...end of the pegRNA, which is not well protected by Cas9. This is similar to the proposed mechanism for engineered...sensors By Brian O’Neill New viral vector preps expressing the Voltron2 voltage indicator, created by the...that administering these AAVs with a dilute Cre-expressing vector resulted in a sparsely-labeled field of...

...involves the expression of long hairpin RNA (hpRNA). This method is facilitated by DNA constructs that enable...research and many different constructs are available to the community. Each construct has unique features, such...transgene. Typically, the desired target of RNAi is expressed as inverted repeats separated by a loop region... such as inducible expression of the hpRNA and/or fusing your gene target sequence to a fluorescent reporter...sequence (discussed below). Examples of these constructs include pGAPi, which allows easy creation of ...reporter. This is achieved at the level of the DNA construct, where your RNAi-target is fused to the DNA backbone...phenotypes quantified. Fluorescent reporter-based RNAi constructs have been designed and widely implemented in ...

...co-conversion markers. Expanding beyond Cas9 Since the development of CRISPR/Cas9 editing methods, other Cas proteins...and yeast cells using modified Cas9 proteins to regulate gene expression (CRISPRi/CRISPRa), visualize specific...University of California, Santa Cruz. Emerging CRISPR/Cas9 editing technologies have transformed the palette...regulation during developmental processes, CRISPR/Cas9 allows us to delete sequences and introduce mutations...remain in the edited animal. Development of CRISPR/Cas9 editing strategies in C. elegans As brilliantly ...step as simple as heat-shocking to induce the expression of a Cre recombinase contained within the knock-in...reported in 2017 uses the direct injection of a CRISPR/Cas9 ribonucleoprotein complex with linear DNA as repair...

...biotin ligases Expression system Stable cell line expressing 1) FLAG-biotin-tagged dCas9, 2) BirA, and ...components are co-expressed, the gRNA targets the dCas9 to the loci of interest and dCas9 is biotinylated...creation of a stable cell line that co-expresses three components: dCas9 with an added biotin acceptor site...cell lines will have varying levels of dCas9 and gRNA expression. Additionally, CAPTURE 3C-based sequencing...that is recognized by biotin ligases normally expressed in eukaryotic cells. Using this tag reduces the...  A second application for CAPTURE 2.0 is the construction of spatial and hierarchical chromatin interaction...conformation data is compared with changes in gene expression, chromatin accessibility, and epigenetic modifications...

...plasmids, catalytically inactive Cas9 (dSpyCas9) fused to APEX2 is expressed from a Tet/Shield1 inducible ...Yale University recently deposited a plasmid for expressing a novel reverse transcriptase (RT), MarathonRT...properties make nanobodies easier to transform and express in bacteria, which, in turn, makes them more adaptable...with biotin as an affinity tag. This ‘Caspex’ construct is guided by a pre-defined sgRNA sequence to a...targeted to a sequence of interest with an sgRNA expressed from a separate plasmid. When cells are treated...genomic loci. He made use of catalytically inactive dCas9 by fusing it to an engineered peroxidase, (APEX)...are enriched in the 20 nm radius surrounding the dSpyCas9-APEX2 fusion.  This Sontheimer lab used this technology...

...Tight control of toxin expression is absolutely necessary, because leaky expression of toxin could select...introduce a mutation in the chromosome is to disrupt expression of a gene with an antibiotic resistance cassette...in the chromosome, sometimes interfering with expression of surrounding genes. However, there are ways...(Thomason et al., 2014). In recent years, CRISPR-Cas9 technology has also aided in the creation of scarless... the second crossover step, they tested their expression under both rhamnose- and arabinose-inducible ...  Giacalone, Matthew J., et al. "Toxic protein expression in Escherichia coli using a rhamnose-based tightly...Valentovich LN. "DNA tuner: a computer program for the construction of polylinker sequences of molecular vectors...

...modulate dCas9 expression, multiple scRNA transcriptional programs can be regulated with dCas9 as a master...) model consists of a catalytically dead Cas9 protein (dCas9) that lacks endonuclease activity coupled...gene expression. This strategy can be applied to effectively change metabolic flux. The dCas9 protein...recruit a transcriptional regulator to modulate expression at that locus. The use of regulators allows a... a greater degree of control of desired expression levels dependent upon the type of transcriptional regulator...are comparatively small allowing multiple sgRNA constructs to be introduced simultaneously in a single plasmid...various transcriptional regulators to alter gene expression at the targeted site. Multiple scaffold RNAs...

... third of the size of Cas9). Figure 2: Comparison of TnpB (A) and Cas9 (B) genome editors. Effector...team characterized and deposited a variety of constructs for visualizing cellular components (nucleus,... mGold2. Top: TIRF-SIM imaging of COS-7 cells expressing LifeAct fused to mVenus, mGold2s, or mGold2t...., recruited by either the ωRNA (TnpB) or a gRNA (Cas9). Drawings not to scale. Created with BioRender.... synapses. In addition, ATLAS itself could be expressed Cre-dependently to allow tracing exclusively from...

...shRNA Expression: Cre-lox can be used to turn shRNA constructs on or off. In floxed-shRNA constructs, Cre...return gene expression to physiological levels. In lox-STOP-lox shRNA constructs, Cre expression promotes...genome or control gene expression or skip ahead to browse highlighted plasmids expressing Cre , Flp , Dre ,...Recognition Sites. Controlling Recombinase Expression By expressing the enzyme at specific times or locations...popular recombinase-dependent gene expression strategies. Gene Expression: A stop codon flanked with loxP...promotes shRNA expression. Gene Switch: These constructs contain two genes of interest, Genes A and B. When ...toxicity. Leakiness Leaky expression, where the gene of interest is expressed even in the absence of the...

... Figure 2: (A) Streamlined VchINT expression constructs. (B) High integration efficiencies achieved...effectors employed by Type II systems (such as CRISPR-Cas9). Unlike in regular Type I CRISPR-Cas systems, however...separate plasmids and multiple promoters were used to express the necessary components in E. coli (Klompe et ...). Recently, the lab introduced a streamlined expression approach (Fig. 2A), involving two-plasmid or ...two-plasmid system, an effector plasmid (pEffector) expresses all seven required genes and the CRISPR RNA under... >99% on-target specificity. Additionally, by expressing multiple crRNAs in parallel (Fig. 3), multiplexed...bacteria, alongside more established tools such as Cas9-recombineering, CRISPRa/i, and base editing. Leveraging...