...guide RNA (sgRNA). While the dCas9 protein provides the transactivation activity, the sgRNA component ...activators and sgRNAs can be separately transfected (pmax dCas9 activators [48223~48227] + sgRNA expression...Activators Gateway Donors ID Plasmid 48214 pAC1-pCR8-dCas9VP48 : dCas9VP48 on Gateway donor vector pCR8/GW... Gateway donor vector pCR8/GW/TOPO. 48219 pAC147-pCR8-dCas9VP64 : dCas9VP64 on Gateway donor vector pCR8...Gateway donor vector pCR8/GW/TOPO. 48221 pAC149-pCR8-dCas9VP160 : dCas9VP160 on Gateway donor vector pCR8...expression vector or the linear DNA containing the sgRNA expressing fragment produced by PCR), or expressed from...(with Stop codon) are also available as Gateway donor vectors [48218~48221] and can be transferred to ...

...using ssDNA oligo as donor template, it's usually more efficient than plasmid donor. Typical design is ...around 400ng of a typical size (~ 5kb) donor plasmid with Cas9/sgRNAs vector, for one well in 24-well plate...around 400ng of a typical size (~5kb) donor plasmid with Cas9/sgRNAs vector, for a one well in a 24-well...notes below are considerations for designing HR donor. Generally if off-target (non-specific) cleavage...then select the best guide to co-transfect in HR donor. Alternatively, a new system described recently ... plate, co-deliver into the cells with the Cas9/sgRNAs vector (like PX330). For large changes (>100bp ...insertions or deletions), we typically use plasmid donor, with two homology arms on each side flanking your...

... Sabatini and Lander gRNA cloning into pLX-sgRNA pLX-sgRNA PDF 125.9 KB Sontheimer gRNA design and cloning...as an optional input to assess HDR frequency. An sgRNA sequence (without PAM sequence) can be provided,... can identify positively or negatively selected sgRNAs, genes, or pathways. CrispyCrunch CrispyCrunch ...CRISPR in 180+ genomes. PubMed PMID: 27380939 . sgRNA Scorer 2.0 From the Church Lab : a tool that identifies... epression, and a ctivation From the Qi Lab , a sgRNA design tool for genome editing, as well as gene ...Cas9 target online predictor Identifies candidate sgRNA target sites by off-target quality. Validated for...Goldstein Nematode: gRNA design and cloning Cas9-sgRNA construct PDF 355.3 KB Goldstein Nematode: Injection...

...an ssODN donor template can mediate gene replacement by homologous recombination. For the sgRNA, vector...51019 pDsRed-attP : Vector for generating dsDNA donors for homology-directed repair to replace genes or.... 51434 pHD-DsRed : vector for generating dsDNA donors for homology-directed repair. Contains the visible...

... targeting the genomic site and the donor vector. The PITCh donor plasmid with an EGFP-2A-PuroR cassette... and donor vectors as well as pX330S-2-PITCh, expressing a generic gRNA targeting the PITCh donor plasmid...expression cassette for the Cas9-programming sgRNA and HR donors for selectable genome modification. This ...Plasmids Protein Species Tag Donor Plasmid gRNA plasmid gRNA plasmid FLAG pFETCh_Donor CEBPB Human FLAG pFETCh_CEBPB...may be used. The HDR (homology directed repair) donor plasmid with homology arms and selection marker....as design and clone in the homology arms for the donor plasmid. The Mendenhall and Myers labs have also...instead of HDR, which makes it easy to construct a donor vector, because MMEJ utilizes very short (around...

...AAV:ITR-U6-sgRNA(Kras)-U6-sgRNA(p53)-U6-sgRNA(Lkb1)-pEFS-Rluc-2A-Cre-shortPA-KrasG12D_HDRdonor-ITR (AAV-...lentiCRISPR - sgRNA and SpCas9 together 2 vector system - lentiCas9-Blast and lentiGuide-Puro (sgRNA) Browse...CMV-driven SaCas9; U6-driven sgRNA 61593 : PX602; TBG-driven SaCas9; U6-driven sgRNA 61592 : PX600; CMV-driven... SaCas9 sgRNA can be co-transfected as PCR amplicons using a protocol similar to SpCas9 sgRNA PCR (Ran... pAAV-U6::sgRNA(SapI)_hSyn-GFP-KASH-bGH (SpGuide acceptor). This is a AAV plasmid for sgRNA cloning. GFP-KASH...implemented by delivering Cre and sgRNA to a Cre-dependent mouse or sgRNA to a constitutively Cas9-expressing...control for AAV-KPL; sgRNA targeted to LacZ, plus luciferase-2A-Cre recombinase 60226 : sgRNA cloning backbone...

...46,354 BARBEKO sgRNA Library 174163 Knockout, Base Editing Human Wei 3rd 3 53,502 sgRNA Pool for Streptococcus...131625 Inhibition E. coli Bikard N/A ∼5 21,417 Bison sgRNA Library 169942 Knockout Human Ebert 3rd 4 2,852 ...Fly Perrimon 3rd 8 6 6 7,956 17,827 59,406 Dual sgRNA CRISPRi Libraries 187246 — 187248 1000000192 Inhibition...Inhibition Human Weissman 3rd 6 Varies Dual sgRNA CRISPRa Libraries 187249 — 187251 1000000193 Activation...3rd 2 37,722 Green monkey (Chlorocebus sabaeus) sgRNA library 178284 Knockout Green Monkey Doench and ...Knockout Human Parts 3rd 3 59,576 Human Whole Genome sgRNA iBAR Library 140633 Knockout Human Wei 3rd 3 58,...HR700_TP53 Exon Mutation Libraries 229137–229140 Donor Vector Human Stiewe N/A Varies 1,910–3,034 per exon...

...specific genomic edits by copying from a DNA donor template . The donor template contains the desired sequence...out integration of donor DNA. Both types of CASTs have been found to achieve donor DNA integration for...systems contain two components: a guide RNA ( gRNA or sgRNA ) and a CRISPR-associated endonuclease (Cas enzyme...cell along with the gRNA(s) and Cas enzyme. The donor DNA contains the desired edit as well as additional...homology arms. Depending on the application, the donor DNA may be a single-stranded oligonucleotide, double-stranded...for large edits or insertions. When designing the donor template, incorporating mutations that prevent further...should alter the PAM site or target sequence in your donor DNA with a silent mutation that does not change ...

...60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR Cre, luciferase, and sgRNA expression EFS... Zhang 60229 AAV:ITR-U6-sgRNA(backbone)-pCBh-Cre-WPRE-hGHpA-ITR Cre and sgRNA coexpression Cbh AAV Zhang...ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR Cre, KASH-tagged EGFP, and sgRNA expression...expression hSyn AAV Zhang 60820 pSECC Cre, Cas9 and sgRNA expression EF-1 alpha Lentiviral Jacks 60877 pMAZe... components, PhyBdeltaCreC and PIF6CreN, in AAV donor/transfer vector Cbh AAV Eisenhoffer 131786 pLenti-CreLite...pAAV-Sox2-CreERT2 Cre-ERT2 - Tamoxifen inducible; AAV donor vector AAV He 135659 pQcre1 Cre PpagA from B. anthracis...

...pyogenes Joung sgRNA with U6 promoter 48962 Worm HindIII none S. pyogenes de Bono sgRNA with rpr-1 promoter...SpBsmBI sgRNA Hygro 62205 Mammalian/Lentiviral BsmBI none S. pyogenes Hygro Maehr pLenti SpBsmBI sgRNA Puro...dCas9-VP64 along with 1-4 sgRNAs expressed from independent promoters. sgRNAs are cloned into separate ... and allow for expression of more sgRNAs in a single vector. sgRNAs are expressed as polycistronic glycine...pLenti-sgRNA-Lib 53121 Mammalian/Lentiviral BsmBI none S. pyogenes ccdB Wei pLKO5.sgRNA.EFS.PAC 57825 ...Church LRG (Lenti_sgRNA_EFS_GFP) 65656 Mammalian/Lentiviral LIC none S. pyogenes GFP Vakoc U6>sgRNA(F+E) 59986... Boxem PU6::klp-12_sgRNA 46170 Worm PCR (see paper) none S. pyogenes Calarco SP6-sgRNA-scaffold 47912 ...

...Walker Jackson 78624 sgRNA(MS2)_Prnp_SAM1 PRNP U6 Dementia Walker Jackson 78625 sgRNA(MS2)_Prnp_SAM2 PRNP...His T5 Parkinson's Suzanne Pfeffer 208392 Lenti-sgRNA-Cre-GpNLuc-sgmATM ATM Ataxia telangiectasia Gaorav...Hemmo Meyer 85845 pDONOR-SNCAe2-WT SNCA CAG Parkinson's Jens Schwamborn 85847 pDONOR-SNCAe3-WT SNCA CAG...Schwamborn 85848 pDONOR-SNCAe3-A53T SNCA CAG Parkinson's Jens Schwamborn 86154 pDONOR-PARK6e5-I368N PINK1...TARDBP-Exon1-ko-sgRNA1-SpCas9 TARDBP U6 ALS Rajat Rohatgi 107858 pHBS953 TARDBP-Exon2-ko-sgRNA2-SpCas9 TARDBP...133762 px459-TDP-43 sgRNA1 TARDBP U6 ALS Shawn Ferguson 133763 px459-TDP 43 sgRNA2 TARDBP U6 ALS Shawn...TIA1 His, T7 T7 ALS Thomas Tuschl 106108 px458_2A_GFP_sgRNA_EIF2AK2 EIF2AK2 U6 VWM disease Thomas Tuschl ...