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Showing: 1 - 20 of 45 results
  1. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    .... U6 small RNA expression shRNA From the human U6 small nuclear promoter Constitutive  Murine U6 is ... trying to pick your perfect promoter! Eukaryotic Promoters   Promoter   Primarily used for RNA     .... In practice, the term "promoter" describes the combination of the promoter (RNA polymerase binding site...recognize and bind to specific promoter elements. This means that the promoter present in your plasmid backbone...RNAP II promoter, whereas small RNAs (such as shRNA) are transcribed from the RNAP III promoters. This ...post on viral vector parts. Promoter specificity Aside from choosing a promoter based on type of RNA transcript...cell types or organisms, promoters must be similarly variable. Bacterial promoters only work in prokaryotic...
  2. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...sequence downstream of the 7SK, human U6, mouse U6, or human H1 promoters. If you express fewer than four gRNAs...from its own promoter. But that doesn’t mean you have to clone and transfect multiple promoter-gRNA constructs... of pX330, contains humanized wtCas9 and two U6 promoters. To use this plasmid, you simply order oligonucleotides...to three gRNAs. Entry vectors containing the U6 promoter and the gRNA scaffold are provided with the kit...cloning. gRNAs are expressed from two Drosophila U6 promoters. Cas9 must be supplied on a separate plasmid...can I express more than one gRNA from a single promoter using a plasmid like pX330? Unfortunately, the...contain Type IIS restriction sites flanking the promoter-gRNA construct, but with different sequences adjacent...
  3. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    ...customizable sgRNA from the U6 promoter and puromycin resistance from the EF-1a promoter. It’s a 3rd generation...the years on their own or through Addgene!) pX330-U6-Chimeric_BB-CBh-hSpCas9 - The S. pyogenes Cas9 (SpCas9...cloned to create a chimeric guide RNA. Find pX330-U6-Chimeric_BB-CBh-hSpCas9. pSpCas9(BB)-2A-Puro (PX459...
  4. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...sequence into a backbone that uses the human U6 promoter to drive expression, is it necessary to add a...the start of my target sequence? A2: The human U6 promoter prefers a 'G' at the transcription start site...
  5. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ...first site accepts the sgRNA target sequence for U6 promoter expression and the second site accepts the homology...containing a direct fusion of the desired gRNA to the U6 promoter, and a second plasmid containing the homology...regions of homology to the gene of interest to promote homology directed repair after a CRISPR/Cas9 induced...
  6. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    ...imporant is that, upstream of the Cbh promoter, this plasmid has a U6 promoter followed by BbsI restriction sites...pyogenes (Sp) Cas9 from a Cbh promoter. Cbh is a modified version of CAG promoter that the Zhang lab has tested...
  7. DAP arrays for multiplex gene editing

    Type
    Blog Post
    ...strategy, the need for lengthy promoters, such as U6 or EF1α promoters, has limited the number of gRNAs...assembled tRNA-gRNAs is placed downstream of a lengthy promoter, which is used to drive the expression of the ...expression of itself, without using any additional promoter, which is followed by tRNA processing to release...human or plant sources) in both the strength as a promoter and the activity to release gRNAs from DAP array...-gRNA arrays, that rely on an upstream lengthy promoter to express the gRNA array, DAP arrays use tRNA...
  8. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...fluorescent proteins, 8 constitutive promoters, 2 includible promoters, 3 polyA terminators and various pieces...sequences. Several pre-constructed promoter-selection marker or promoter-inducible expression related vectors...all chemically inducible promoters; 3) improving the strength of the promoters used; and 4) optimizing ...ideal promoter/location combination. However, other constructs which contain different promoters and/or...to study the human GM-CSF promoter and enhancer, a finely regulated promoter controlled by a mixture of...containing a promoter for constitutive (CMV, SV40, SFFV) or inducible (CMV-TO) cDNA expression, sgRNA (U6) expression...the available sgRNA-cargo constructs utilize a U6 promoter and contain the RNA cargo cassette inserted within...
  9. Hot Plasmids - October 2020

    Type
    Blog Post
    ...express hybrid Cas9-Cas12a gRNAs under a single U6 promoter. The paralog and dual-targeting hybrid gRNA library...
  10. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...Corn under promoters for transcripts uniquely transcribed by RNA Polymerase III (5S, tRNA and U6) and measuring...transcription by placing the aptamer downstream of a known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores ...
  11. Qi Lab CRISPR Page

    Type
    Collection
    ...pSico-based U6 vector containing murine U6 promoter and sgRNA targeting GAL4 UAS promoter 46916 pU6-sgGAL4...pSico-based U6 vector containing murine U6 promoter and sgRNA targeting GAL4 UAS promoter (negative control...retroVirus LTR promoter. The second plasmid ( pgRNA_humanized ) contains a murine U6 promoter controlled ...pU6-sgGFP-NT1 Human pSico-based U6 vector containing murine U6 promoter and sgRNA targeting GFP (NT1) 46917...pU6-sgCXCR4-2 Human pSico-based U6 vector containing murine U6 promoter and sgRNA targeting endogenous ...pU6-sgCD71-2 Human pSico-based U6 vector containing murine U6 promoter and sgRNA targeting endogenous ...contains sgRNA controlled by SNR 52 promoter, targeting endogenous TEF1 promoter 46923 pSNR52-sgTET Yeast CEN...
  12. CRISPR Fly Design - Drosophila genome engineering

    Type
    Collection
    ... under control of the Drosophila U6:3 promoter Bullock 49411 pCFD4-U6:1_U6:3tandemgRNAs expression of ...expression of gRNA under control of the Drosophila U6:1 promoter Bullock 49409 pCFD2-dU6:2gRNA expression of ...of gRNA under control of the Drosophila U6:2 promoter Bullock 49410 pCFD3-dU6:3gRNA expression of gRNA ...of two gRNAs from Drosophila U6:1 and U6:3 Bullock 73914 pCFD5 multiplex gRNA plasmid Bullock 73915 pCFD6...pAct-Cas9 Expresses Cas9 under control of act5C promoter and SV40 3'UTR. For ubiquitous genome engineering...pnos-Cas9-nos Expresses Cas9 under control of nanos promoter and 3'UTR. For germ line restricted genome engineering...dCas9 Expresses Fok1:dCas9 under control of act5C promoter and SV40 3'UTR. For ubiquitous high specificity...
  13. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ...Cas9T2AGFP;U6:sgRNA1;U6:sgRNA2 74009 Zebrafish U6 yes, cut S. pyogenes EGFP Del Bene pUAS:Cas9T2ACre;U6:sgRNA1...sgRNA with U6 promoter 48962 Worm HindIII none S. pyogenes de Bono sgRNA with rpr-1 promoter 48961 Worm...of two sgRNAs from independent U6 promoters and Cas9 from CBh promoter. Ventura A CRISPR/Cas9 toolkit ...Uses five distinct zebrafish U6 promoters for sgRNA expression. Chen pCFD4-U6:1_U6:3tandemgRNAs Drosophila...expression of two gRNAs from Drosophila U6:1 and U6:3 promoters. Bullock pCFD5 Drosophila Utilizes endogenous... Marraffini pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) 47868 Mammalian U6 yes, cut N. meningitidis...LEU2 Lu pDestTol2CG2-U6:gRNA 63156 Zebrafish BseRI none S. pyogenes Zon pDestTol2pA2-U6:gRNA 63157 Zebrafish...
  14. CRISPR Plasmids - Plants

    Type
    Collection
    ...pHAtC U6 AarI yes, cut S. pyogenes Hygro Kim pBAtC U6 AarI yes, cut S. pyogenes Basta Kim pHEE401 U6-26p...pyogenes Gao pU6-gRNA wheat U6 BbsI none S. pyogenes Gao pBlu/gRNA Arabidopsis U6 BbsI none S. pyogenes Stupar... Chen pHEE401E U6-26p BsaI yes, cut S. pyogenes Hygro Chen pHDE-35S-Cas9-mCherry-UBQ U6-26 PmeI yes, cut...opposite strand) mutations. Plasmid Gene/Insert Promoter Selectable Marker PI Publication Nick CRISPR/Cas...homology-directed repair (HDR). Plasmid Gene/Insert Promoter Selectable Marker PI Publication Prime Edit Cas9...desired edits on an RT template. Plasmid Gene/Insert Promoter Selectable Marker PI Publication Activate Catalytically...gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene of interest...
  15. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ....org/10878/ . Description Vector Element U6 Human U6 promoter drives RNA Polymerase III transcription ...transduced cells. hPGK Human phosphoglycerate kinase promoter drives expression of puromycin. Puro R Puromycin...
  16. Kamoun Lab CRISPR Plasmids

    Type
    Collection
    ... 35S promoter in the plant tissue. 46968 pICSL01009::AtU6p : Encodes the Arabidopsis U6 promoter in a ... used to place an sgRNA uder the Arabidopsis U6 promoter. 46966 pICH86966::AtU6p::sgRNA_PDS : Expresses...in Nicotiana benthamiana from the Arabidopsis U6 promoter. 48075 pICH86966 : A binary vector for plant ...
  17. CRISPR Plasmids - Prime Edit

    Type
    Collection
    ...Gene/Insert Promoter Selectable Marker PI Publication Bacteria Plasmid Gene/Insert Promoter Selectable ...Gene/Insert Promoter Selectable Marker PI Publication Drosophila Plasmid Gene/Insert Promoter Selectable...Drosophila U6:3 pegRNA BbsI No Vermilion Norbert Perrimon pCFD5-NS Drosophila Drosophila U6:3 pegRNA +...find a gRNA vector based on expression system, promoter, the type of gRNA (e.g., pegRNA, epegRNA, nicking...plasmid contains Cas9. Plasmid Expression System Promoter Guide RNA Type Cloning Enzyme Co-expressed Cas9...BbsI No Vermilion Norbert Perrimon pHSG1C3 Mammalian U6 pegRNA + nicking sgRNA BbsI for sgRNAs; BbsI + PstI...tmpknot-GG-acceptor Mammalian hU6 epegRNA BsaI No mRFP1 David Liu U6-pegRNA-H1-nick sgRNA-mCherry Mammalian, AAV hU6 +...
Showing: 1 - 20 of 45 results