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  1. Zhang Lab's CRISPR Frequently Asked Questions

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    ...use double nickase, you could express two spacers and use PX335 to express the Cas9n (nickase). The concept...oligo when cloning my target spacers into the PX330 vector backbone? There is no need to add the NGG PAM...start site to have high expression, so adding this G could help with expression, though it is possible ...you need to have four spacers co-expressed and using PX335 to express the Cas9n (nickase). Homologous ...co-deliver into the cells with the Cas9/sgRNAs vector (like PX330). For large changes (>100bp insertions or...the second design (in pX330 / PX335 ), so we recommend people all use the pX330/PX335 system ....concept of the double nickase system is that you can express two different chimeric gRNAs with the Cas9 nickase...

  2. CRISPR Plasmids - Tagging

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    ...Tagging Plasmids: pX330S-2-PITCh - expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh - expresses Cas9, PITCh-gRNA...gene-specific CRISPR and donor vectors as well as pX330S-2-PITCh, expressing a generic gRNA targeting the...targeting vector assembly, and the remaining 5 plasmids include Cre and FLP expression vectors, a general... one vector from the Doyon lab, eSpCas9(1.1)_No_FLAG_AAVS1_T2 (Addgene #79888) , which expresses an untagged...components: An all-in-one CRISPR/Cas9 vector based on the Zhang lab's PX330 , simultaneously targeting the genomic...are available. An improved FLP recombinase expression vector allows for efficient marker cassette FLP-out...-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila ...

  3. Genomic Deletions in Mammalian Cell Lines

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    ...the 20-mer is not G. sgRNA expression from the U6 promoter of the pX330 vector is enhanced by the inclusion...cloning and expression purposes. NOTE: Here we discuss the usage of the pSpCas9(BB) plasmid (pX330) (Addgene...guide’s reverse complement for cloning into the pX330 vector using BbsI restriction enzyme ( Table 3 ). Add...Prepare a 50 μl reaction mix: 100 ng circular pX330 vector, 1.0 μl annealed oligos (1 μM; see step 3.1.4...42230). This plasmid allows for the simultaneous expression of sgRNA and SpCas9, but does not contain markers...or constructs in which multiple sgRNAs may be expressed from a single plasmid. Add “CACC” before the 20...concentration of 1 μM). Ligate annealed oligos into pX330 using a Golden Gate assembly cloning strategy 3 ...

  4. CRISPR References and Information

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    ...cloning vector Retroviral vectors: neomycin (pSIR-neo) , GFP (pSIR-GFP) , DsRed (pSIR-DsRed-Express2) , human... sgRNA dual expression: pAC2 , pAC152 , pAC153 , pAC154 ; pmax dCas9 activator expression: pAC91 , pAC92...Lines pSpCas9(BB) (pX330) Protocol at Addgene Parrott NEBuilder Assembly of CRISPR vectors using ssDNA oligos... pooled library amplification 1 vector system: lentiCRISPR v2 2 vector system: lentiCas9-Blast and lentiGuide-Puro...Addgene Church gRNA design and cloning gRNA cloning vector PDF, 118 KB Church gRNA design and cloning for ...human pluripotent stem cells pCas9_GFP ; gRNA empty vector Protocol at StemBook O'Connor-Giles Fly: gRNA and...design and cloning All-in-one plasmid containing expression cassette for NmCas9 and BsmBI site for cloning...

  5. CRISPR Plasmids - Empty gRNA Vectors

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    ... Empty Vectors Many gRNA empty vectors have been deposited at Addgene. To find the gRNA vector for your...constructing all-in-one expression vectors containing multiple guide RNA expression cassettes and a Cas9 ...size and allow for expression of more sgRNAs in a single vector. sgRNAs are expressed as polycistronic ...gRNA expression vectors for insertion of custom gRNA target sequences. CRISPR...CRISPR Plasmids: Empty gRNA Expression Vectors Browse CRISPR Plasmids By Function Genome Editing Cut Base...by the Doudna Lab . Plasmid Expression System Cloning Enzyme Co-expressed Cas9 Cas9 System Selection PI...BbsI none S. pyogenes Gersbach pUC57-sgRNA expression vector 51132 Mammalian BsaI none S. pyogenes Huang...

  6. Zhang Lab CRISPR Page

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    ...2015). The dual vector system uses S. pyogenes Cas9 (SpCas9), using one vector to express SpCas9, and another... in two formats: 1 vector system - lentiCRISPR - sgRNA and SpCas9 together 2 vector system - lentiCas9...transfection : 61422 : Expresses dCAS9-VP64 activator with 2A GFP 61423 : Expresses the MS2-P65-HSF1 activator...lentiviral vector encoding dCAS9-VP64 with 2A Blast resistance marker 61426 : lentiviral vector encoding...sgRNA to a constitutively Cas9-expressing mouse. Described here are AAV vectors that can be combined with ...61408 : Targeting vector for the mouse Rosa26 locus; Used to make Cas9 knockin mouse Vectors for cancer modeling...toolbox protocol 637.8 KB CRISPR-Cas9 Cre expression vectors for cancer modeling Using Cas9 mice, Platt...
Showing: 1 - 6 of 6 results