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We narrowed to 89 results for: grna cloning vector

Showing: 41 - 60 of 89 results

  1. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    ...the ITRs does not get packaged in the AAV vector. The cloning capacity between the ITRs is ~4.4 kb. ITRs...smaller CRISPR-Cas proteins such as SaCas9, CRISPR gRNAs, or your gene of interest. Promoter The promoter... the Addgene blog Learn more about basic viral vector elements Read about four considerations for the...

  2. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...consists of highly useful vectors compatible with Gibson and Saptrap cloning systems to generate AID* ... about a week. Taking advantage of Golden Gate cloning, the Haynes lab’s Protocols.io protocol describes...for delivering CRISPRi components by a single AAV vector are now available from Tonia Rex's lab.  vfCRISPR... system developed by Taekjip Ha’s lab where Cas9-gRNA complex binds the target DNA without cleavage until... Gether lab, now available as ready-to-use AAV vectors.   ...

  3. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    ...suited (Gaudelli et al., 2017). Our vectors have two Bsa1 sites to clone the single guide RNA (protospacer...We use a single polycistronic tRNA-gRNA (PTG) gene system to clone multiple protospacers which enables...plant We developed plant adenine base editor (ABE) vectors to install A>G or T>C mutations in the plant genome...genome. We constructed three versions of the vector (pKABE6.3, pKABE7.9, and pKABE7.10) so that the user... genomic site simultaneously.     These binary vectors could be used to generate stable genome edited ...Agrobacterium-mediated transformation. We also constructed smaller vectors suitable for transient assays using protoplast ...

  4. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ...you have one AAV vector dedicated to expressing Cas9, while the other houses the gRNA and scaffold. If...packaging capacity and stable expression of lentiviral vectors make them an attractive option. However, safety... particularly in vivo ones. In those cases, AAV vector screens may be AAVantageous!       In vivo, in ...delivery method for in vivo CRISPR screens. AAV vectors can be made at high titers equal to those achieved... customization that is unmatched by other viral vector delivery methods. Lentivirus has limited packaging...packaging options, with most lentiviral vectors being packaged using VSV-G. In contrast, AAV can be packaged...psuedotypes to further improve the tropism of AAV vectors, enhancing their affinity for specific cell and...

  5. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT vector ensures...desired insert is cloned using traditional restriction enzyme cloning to create an Entry vector containing a...with custom-cloned vectors in a Multisite Gateway reaction with a desired destination vector expressing...standard entry clone. ORFs from an ORFeome Gateway library or any PCR-amplified ORF cloned into pDONR221... promoter-less Gateway® entry clones to be used with other entry clones encoding the ORFs of interest:...science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with ...building-block vectors that are combined in a specified order using Multisite Gateway cloning. A desired ...

  6. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...Addgene, including cDNA, barcoding, screening, and gRNA libraries. Most of the pooled libraries at Addgene...protocols and guides regarding pooled libraries, from cloning a pooled library to viral production to screening...

  7. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...genomic targets. These two vectors are part of the larger pMVP/pMAGIC cloning system, which contains Cas9...methylation and activate transcription. A lentiviral vector with the same modifier, Fuw-dCas9-Tet1CD, is available... targeted enhancers. Although the lab used this vector in chick embryos, it also functions in mammalian...and this fusion allows for packaging into viral vectors and targeted delivery into hard-to-manipulate cell...Arlotto ME, et al (2018) Creation of versatile cloning platforms for transgene expression and dCas9-based...Check out Our CRISPR Guide Pages Find Validated gRNAs to Target Your Genomic Locus of Interest Browse ...

  8. CRISPR References and Information

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    Collection
    ... Church gRNA design and cloning gRNA cloning vector PDF, 118 KB Church gRNA design and cloning for Cas9... PDF, 66.8 KB Fujii gRNA design and cloning gRNA cloning vector Retroviral vectors: neomycin (pSIR-neo...and Lander gRNA cloning into pLX-sgRNA pLX-sgRNA PDF, 129 KB Sontheimer gRNA design and cloning All-in-one...Cas9 and gRNA use Cas9 (pIK86) ; gRNA empty backbone (pDR274) PDF, 89 KB Liu Fly: gRNA cloning pAc-sgRNA-Cas9...GFP ; gRNA empty vector Protocol at StemBook O'Connor-Giles Fly: gRNA and ssODNs design and cloning; injection...Chen and Wente Zebrafish: gRNA cloning, in vitro transcription, injection gRNA core ; Cas9 ; optimized ...dCas9 PDF, 110 KB Goldstein Nematode: gRNA design and cloning Cas9-sgRNA construct PDF, 364 KB Goldstein Nematode...

  9. CRISPR Plasmids - Tagging

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    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...components: A vector containing Cas9 and a validated gRNA, based on the Zhang lab's PX458 . Multiple gRNA plasmids...expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh - expresses Cas9, PITCh-gRNA, & FBL-specific gRNA pCRIS-PITChv2-FBL...currently unavailable, you will need to design and clone in a gRNA(s) to guide the Cas9 protein to your target...Plasmids Protein Species Tag Donor Plasmid gRNA plasmid gRNA plasmid FLAG pFETCh_Donor CEBPB Human FLAG...of native protein complexes. This vector serves as a backbone to clone the left and right homology arms...2015 . Alternatively, cDNAs can be cloned directly into this vector and targeted to the AAVS1 genomic ...

  10. CRISPR Plasmids - Empty gRNA Vectors

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    Collection
    ...genome. gRNA Empty Vectors Many gRNA empty vectors have been deposited at Addgene. To find the gRNA vector... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Multiplex gRNA Vectors Several systems have been developed to generate multiplex gRNA vectors. These systems...single gRNA vectors with various promoters, plasmids for assembling multiple gRNAs into one vector, and ...Empty gRNA expression vectors for insertion of custom gRNA target sequences. CRISPR...U6 yes, cut (enhanced) S. pyogenes Zhang gRNA Cloning Vector Bbs I ver. 2 85586 Mammalian BbsI None S....designed for monocots. Using these gRNA module vectors, multiple gRNA expression cassettes can be assembled...

  11. Plan Your Experiment

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    Collection
    ...the edit type, designing and cloning a gRNA, delivering Cas and the gRNA, and validating the edit....more about how to design your gRNA . Synthesize and Clone Desired gRNAs Once your target sequences are...it’s time to design your gRNA oligos and clone them into your desired vector. In many cases, you can synthesize...containing the gRNA scaffold using standard restriction-ligation cloning. However, the exact cloning strategy...about viral vectors and their production, see our viral vector guides . Lentiviral vectors Lentiviral ...lentiviral preps . AAV vectors AAV vectors offer some advantages for viral vector delivery. Both lentiviral...Introduction Select Genetic Manipulation Design and Clone gRNA Select Expression and Delivery Validate Genetic...

  12. CRISPR Plasmids - Zebrafish

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    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Gene/Insert Vector Type Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression ...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR...double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially...flanking the DSB and a specific edit close to the gRNA PAM site. When a repair template is present, the...

  13. CRISPR Plasmids - Xenopus

    Type
    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR...) at a specific location based on a co-expressed gRNA-defined target sequence. DSBs are preferentially...surrounding the DSB is introduced along with the Cas9 and gRNA plasmids, the cell may instead repair the break ...

  14. Plant Plasmids and Resources

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    Collection
    ... maize and Arabidopsis) and vectors for easy assembly of one or more gRNA expression cassettes - Qi-Jun...Caixia Gao. Cloning The following collections are designed for creating plant expression vectors: A modular...Martin Parniske. GreenGate Cloning System to create plant expression vectors containing several cassettes...Scott Lenaghan. Joint Modular Cloning (JMC) Toolkit includes a modular cloning chassis and set of functional...microRNA and siRNA vectors in Arabidopsis - James Carrington. JoinTRV , an improved vector system based on...as the promoter or cloning method of choice. ID Plasmid Description Promoter Cloning Method PI Return to...Protein Guide Addgene Blog: Plant Biology CRISPR Cloning Transcription Factors RNAi Empty Backbones Fluorescent...

  15. CRISPR Plasmids - C. elegans

    Type
    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR...double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially...

  16. CRISPR Plasmids - Drosophila

    Type
    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR...double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially...

  17. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ...Addgene blog Viral Vector Elements The Retroviral Life Cycle Popular Retroviral Vectors and Their Use in...  Frequently asked questions about γ-retroviral vectors How do engineered γ-retroviruses differ from lentiviruses...example: pLXSN has the wild type LTR, an MCS for cloning X gene, an SV40 promoter, and Neomycin selection...either side of a retroviral provirus (see below). Cloning capacity between the LTRs is ∼8.5 kb, but inserts...was originally published in 2018, in the Viral Vectors 101 eBook, 1st edition.  References and resources...γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA of interest: The transgene sequence is...

  18. CRISPR Plasmids - Yeast

    Type
    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Selectable Marker PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...using does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Validated In Resistance Co-expressed...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR...

  19. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...single cell clones for in vitro CRISPR experiments. The likelihood that a single cell clone derived from...position of the mismatch along the length of the gRNA given the stricter sequence requirement at the terminal...may not be worth the cost and labor when single clones are selected. Conversely, translational research...and TALENs using integrase-defective lentiviral vectors." Nature biotechnology 33.2 (2015): 175-178. PubMed...many software tools predict off-target effects of sgRNAs using computational methods. They identify possible...mismatches based on the sequences of the genome and sgRNA. This is a good starting point for most researchers...

  20. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...fusions for membrane display, they recommend the NGR vectors for most experiments. These new tools promise to... toolkit is the empty plasmid backbone: the DNA vector that contains everything needed to replicate inside...backbone plasmids here! Figure 2: Addgene cloned EGFP into pAG Lenti CMV N-HA Puro (Addgene #236079...scarless insertion of the payload (teal). The Cas9 gRNA targeting guide and the transposon RNA homology ...successful experiments or therapies. Using AAV vectors with enhancers that drive cell-type-specific expression...colleagues characterized a series of new enhancer-AAV vectors targeting specific classes of cortical and striatal...
Showing: 41 - 60 of 89 results