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We narrowed to 55 results for: dcas9 coding sequence

Showing: 21 - 40 of 55 results

  1. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...easy enough to contemplate DNA in terms of its sequencing, in vivo studies should always take DNA’s chromatin... editor targets DNA (via catalytically inactive dCas9) to modify chromatin. A fluorescent protein can ...expression pattern and an auxin inducible degron (AID) sequence attached to your protein of interest. TIR1 interacts...of auxin, this TIR1 complex binds to the degron sequence ultimately leading to the degradation of the AID-tagged...(minimal 44 amino acids auxin inducible degron sequence) tagged proteins. Degradation of your protein ...Jacobs-Wagner’s lab produce different levels of dCas9 expression and contain different antibiotic resistance...Kleinstiver’s labs are now available. These libraries encode target sites flanked by randomized nucleotides ...

  2. Brzezinski Lab CRISPR Collection

    Type
    Collection
    ... 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...

  3. Bikard Lab - CRISPR Repression Collection

    Type
    Collection
    ... 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...

  4. CRISPR Plasmids - Bacteria

    Type
    Collection
    ...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 ...

  5. CRISPR Plasmids - Yeast

    Type
    Collection
    ...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 ...

  6. CRISPR Plasmids - Mammalian Expression

    Type
    Collection
    ...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...

  7. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...antibiotic resistance selection markers, gRNAs, and dCas9 proteins. Second, it can be introduced into a broader...CRISPRi cassette flanked by left and right Tn7 end sequences. Donor bacteria with a chromosomal copy of the...recipient bacteria due to site selection by TnsD, a sequence-specific DNA binding protein. Selection removes...DNA (purple) flanked by left and right Tn7 end sequences (green) integrates downstream of the glmS gene...promoters, selection markers, and different versions of dCas9. Following integration, Mobile-CRISPRi is stably...s take a look at the essential gene folA which encodes the target for the antibiotic trimethoprim. When...

  8. CRISPR Plasmids - Repress Gene Expression

    Type
    Collection
    ...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...

  9. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...reconstituting coding sequences for a range of applications, including CRISPR activation by dCas9-VPR, prime...platform is compatible with enrichment, single-cell sequencing, and optical pooled screens in human cells. Addgene... improve the specificity of ssAAV, a targeting sequence of miR124T was inserted into the vector to silence...alone or in combination with miR124T targeting sequences, provides a simple yet valuable tool for microglia...find in this post: Multiplexed perturbation and decoding in pooled CRISPR screens Improved prime editor...Collection updates   Multiplexed perturbation and decoding in pooled CRISPR screens By Emily Bentley Building...1: CROPseq-multi uses two sgRNAs with internal barcodes (iBARs), multiplexed using tRNAs, within the lentiviral...

  10. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ... The popular CRISPR base editing technique uses dCas9/Cas9 nickase fused to a cytidine deaminase for targeted...develop five new base editors with distinct PAM sequences, expanding the number of available target sites...Knockdown library. Although B. subtilis has a fully sequenced genome, many applications in biotech, and is considered...non-essential. The knockouts replace entire protein-coding regions (excluding start and stop codons) with ...Each individual strain includes a xylose-inducible dCas9 paired with a single, unique, and constitutively... applications, such as reversing the effects of dCas9 binding to a genomic locus, or limiting the amount...physiology and molecular processes via genetically encoded proteins. PhoCl is a new member of this class that...

  11. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...which consist of plasmids that over-express coding sequences from a given cell type or organism, have been...environmental triggers. This is done by creating a split dCas9 protein that can reassemble when exposed to specific... however, Cas9 allows you to easily change the sequence targeted by the activator by simply providing ...several popular CRISPR-based tools for altering DNA sequence or regulating gene expression.     References ...

  12. Plan Your Experiment

    Type
    Collection
    ...genome sequence dCas9 or dCas9 fusion with transcriptional repressor (such as dCas9-KRAB) dCas9-KRAB can...modifying the genome sequence dCas9 fusion with transcriptional activator (such as dCas9-VP64) Many different...perfect world, your gRNA sequence would be an exact complement to your target sequence with no homologous sites... the desired target sequence, or on-target activity. Two gRNA targeting sequences with 100% homology to... selected target sequence. Additionally, any mismatches between a given gRNA sequence and your genomic...customization Usually paired with a sequencing method Can’t provide specific sequence information alone Digital ...rare edits Many options like RNA-sequencing, Perturb-seq, deep sequencing, etc. qPCR Knockouts Activation...

  13. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    ...proteins, dCas9, and a modified gRNA to direct a variety of customizable functions to your DNA sequence of interest...technology that can be used to quickly barcode a set of DNA sequences. In this post, Alan Wong (formerly ...described in this post are engineered to bind RNA sequences derived from Zika virus and, upon binding, produce...stripped down viral genomes containing only the sequences required for viral replication and transcription... Lab at MIT) shows how you can use CombiGEM to barcode small libraries of CRISPR gRNAs to use for screening...

  14. CRISPR Plasmids - Plants

    Type
    Collection
    ...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...

  15. CRISPR Plasmids - Parasites

    Type
    Collection
    ...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...

  16. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...DNA containing basic parts (promoters, UTRs, coding sequences, terminators, etc) into individual Level 0...insert modules (plant promoter, N-terminal tag, coding sequence of the gene of interest, C-terminal tag, plant...Timer protein fused to the mitochondrial targeting sequence of cytochrome c oxidase subunit VIII, so that ...exchange and re-use of existing elements. DNA sequences for a desired component are cloned into Golden...channelrhodopsins – Chronos & Chrimson Through de novo sequencing of 127 algal transcriptomes, as well as further...for CRISPR gRNA validation Selecting a gRNA sequence that effectively targets your gene/region of interest...their gRNAs. The first step is to clone the target sequence in between two fragments of EGFP in pCAG-EGxxFP...

  17. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

    Type
    Blog Post
    ...appropriate promoter sequence and activation of gene expression. In addition to plasmids encoding these synthetic...CRISPR and RNAi have helped researchers alter DNA sequence and RNA expression - but what if you just want...modulation. While catalytically inactive cas9 (e.g. dCas9) has been fused to a variety of protein modules ...from the El-Samad lab recently deposited plasmids encoding synthetic yeast transcription factors that can...factors, the El-Samad lab has deposited plasmids encoding fluorescent reporters and other proteins controlled...

  18. Zhang Lab CRISPR Page

    Type
    Collection
    ...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...

  19. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    ...about CASANOVA? It works with the S. pyogenes Cas9, dCas9-effector fusions, and xCas9, and requires no additional... mVenus linked to a mitochondrial localization sequence TOM20 (pTriEX Tom20 mVenus BoPD). In dark adapted...plasmids encoding a fusion between cPYP and tgRFP (PLL7.0 tgRFP cPYP), and plasmids that encode the protein...expensive. Thus scientists have turned to genetically encoded voltage indicators. To expand the voltage indicator...

  20. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

    Type
    Blog Post
    ...components: A plasmid in the host bacterium expressing dCas9 A plasmid in the host bacterium that expresses gene...editors capable of editing previously uneditable sequences. Four DNA base swaps, so called point mutations...corrects early stop codons within T7 RNAP. A phage encoding the deaminase TadA-7.10 A mutagenesis plasmid ...
Showing: 21 - 40 of 55 results