...cloning gRNA cloning vector Retroviral vectors: neomycin (pSIR-neo) , GFP (pSIR-GFP) , DsRed (pSIR-DsRed-Express2...pAc-sgRNA-Cas9 PDF 165.1 KB Marraffini Bacteria: pCas9 new spacer cloning pCas9 PDF 125.6 KB Marraffini Bacteria: pCRISPR...CRISPRs in human pluripotent stem cells pCas9_GFP ; gRNA empty vector Link O'Connor-Giles Fly: gRNA and ssODNs...Mammalian Cell Lines pSpCas9(BB) (pX330) Link Parrott NEBuilder Assembly of CRISPR vectors using ssDNA oligos...positive control: CMV-EGFP PDF 2.3 MB Zhang GeCKO pooled library amplification 1 vector system: lentiCRISPR...protocol Church gRNA design and cloning gRNA cloning vector PDF 115.2 KB Church gRNA design and cloning for...lentiCRISPR v2 2 vector system: lentiCas9-Blast and lentiGuide-Puro packaging plasmids: pVSVg , psPAX2 positive...

...expression vector containing N-terminal, TEV cleavable, 6x-His and thioredoxin tags. Other empty vectors for ...many different restriction enzymes. pPro24-gfp contains GFP but all others are empty. pdCas9-bacteria ...Various Plasmids pTac GFP, Luciferase (LuxCDABE) Attila Karsi Broad host range vectors for expressing luciferase... your research. Plasmid ID Technique PI Purpose pCas9 42876 CRISPR Luciano Marraffini Bacterial expression...PIs Fluorescent protein fusion: contains empty vectors with N and C-terminal fluorescent protein tags .... pCyPet-His 14030 FRET Patrick Daugherty FRET: Vector for arabinose inducible expression of an N-terminally... . pYPet-His 14031 FRET Patrick Daugherty FRET: Vector for arabinose inducible expression of an N-terminally...

... Empty Vectors Many gRNA empty vectors have been deposited at Addgene. To find the gRNA vector for your...Kim pCAG-SpCas9-GFP-U6-gRNA 79144 Mammalian hU6 yes, cut S. pyogenes EGFP Zou pCAG-eCas9-GFP-U6-gRNA 79145...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 plasmids...Cerulean Church LRG (Lenti_sgRNA_EFS_GFP) 65656 Mammalian/Lentiviral LIC none S. pyogenes GFP Vakoc U6>sgRNA...cut Lb Cpf1 Neo Welker AIO-GFP 74119 Mammalian U6x2 yes, nick S. pyogenes EGFP Jackson AIO-mCherry 74120...expression vectors for insertion of custom gRNA target sequences. CRISPR...

.... 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...activator with 2A GFP 61423 : Expresses the MS2-P65-HSF1 activator helper complex with 2A GFP 61424 : sgRNA...lentiviral vector encoding dCAS9-VP64 with 2A Blast resistance marker 61426 : lentiviral vector encoding...(SapI)_hSyn-GFP-KASH-bGH (SpGuide acceptor). This is a AAV plasmid for sgRNA cloning. GFP-KASH fusion ...61408 : Targeting vector for the mouse Rosa26 locus; Used to make Cas9 knockin mouse Vectors for cancer modeling... human codon-optimized SpCas9 or SpCas9n, and the single guide RNA. The vector can be digested using BbsI...

...17446 pLenti CMV GFP Hygro (656-4) GFP Hygromycin 3rd gen lentiviral eGFP expression vector, CMV promoter...more details about our viral vector packaging service, visit our Viral Vector Packaging Service page . Pooled... 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP Zhang 61425 ...Ready-to-use lentivirus available from Addgene's viral vector packaging service. Browse constructs currently ... Viral Vector Packaging Service Lentivirus Browse In-Stock Lentivirus... (D10A, N863A) Blasticidin 3rd generation lenti vector encoding dCAS9-VP64 with 2A Blast resistance marker...MS2-P65-HSF1_Hygro MS2-P65-HSF1 Hygromycin lenti vector encoding the MS2-P65-HSF1 activator helper complex...

...targeting vector assembly, and the remaining 5 plasmids include Cre and FLP expression vectors, a general... cloning plasmid, and a prebuilt Unc-32::GFP targeting vector. Jorgensen Lab SapTrap CRISPR/Cas Toolkit... into this vector and targeted to the AAVS1 genomic safe harbor locus using untagged SpCas9 (Addgene 41815...Addgene #41818) or using an all in one vector from the Doyon lab, eSpCas9(1.1)_No_FLAG_AAVS1_T2 (Addgene #79888...the genomic site and the donor vector. The PITCh donor plasmid with an EGFP-2A-PuroR cassette, flanked by...FBL-specific gRNA pCRIS-PITChv2-FBL - PITCh donor vector for EGFP-2A-PuroR knock-in into human FBL locus Jorgensen...-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila ...

... PINK1 N-GFP PINK1 GFP CMV Parkinson's Mark Cookson 13316 pcDNA-DEST47 PINK1 C-GFP PINK1 GFP CMV Parkinson's...21190 GFP-pcDNA3-PKCgamma-cys1Acys1B PRKCG GFP CMV Spinocerebellar ataxia 27 Tobias Meyer 21204 GFP-N2-PKCgamma...PKCgamma PRKCG GFP CMV Spinocerebellar ataxia 26 Tobias Meyer 21205 GFP-C1-PKCgamma-C1A PRKCG GFP CMV Spinocerebellar...-HtrA2-EGFP HTRA2 GFP CMV Parkinson's L. Miguel Martins 14122 pcDNA3-HtrA2-EGFP S306A HTRA2 GFP CMV Parkinson's... GPD 25QDProGFP p416 HTT GFP GPD Huntington's Susan Lindquist 15569 GPD 104QDProGFP p416 HTT GFP GPD Huntington's...GAL 25Q+ProGFPp416 HTT GFP GAL1 Huntington's Susan Lindquist 15581 GAL 46Q+ProGFPp416 HTT GFP GAL1 Huntington's...GAL 72Q+ProGFPp416 HTT GFP GAL1 Huntington's Susan Lindquist 15583 GAL 103Q+ProGFPp416 HTT GFP GAL1 Huntington's...

... pX459 (Addgene plasmid ID 48139), which include GFP and puromycin as selectable markers, respectively...s reverse complement for cloning into the pX330 vector using BbsI restriction enzyme ( Table 3 ). Add ...sgRNA expression from the U6 promoter of the pX330 vector is enhanced by the inclusion of a G nucleotide ...Prepare a 50 μl reaction mix: 100 ng circular pX330 vector, 1.0 μl annealed oligos (1 μM; see step 3.1.4 )...CRISPR/Cas9 construct (10 μg total). Add 0.5 μg of GFP expression construct. Electroporate cells with 250...filter into a FACS tube. FACS sort the top ~3% of GFP positive cells in order to enrich for cells that ... ( e.g., with a PCR cloning kit) into a plasmid vector. Transform the plasmid into DH5α E. coli cells ...

...format into lentiviral transfer vectors (Figure 8B). Libraries come in 1-vector systems, in which Cas9 is included...targeting in the double digits. These multiplex gRNA vectors can be used to knock out, activate, or repress ...multiplexing . Browse Plasmids: Multiplex gRNA Vectors Engineering the Cas9 Endonuclease CRISPR specificity...heterogeneous population of lentiviral transfer vectors. In most cases, each plasmid contains an individual... included in the gRNA-containing plasmid, or 2-vector systems, in which Cas9 must be delivered separately...experimental cell population (Figure 8E). In a 2-vector system, you’ll need to either co-infect with a ...fluorescent marker like green fluorescent protein (GFP), creating a customizable DNA or RNA label for fluorescence...

...and Cas9 expression vectors and validated the use of RNAs produced by these vectors for efficiently modifying...oligonucleotides needed to construct gRNA expression vectors. More information about RGNs and CRISPR-Cas systems... (AsCpf1 and LbCpf1) for use in mammalian cells SpCas9 variants with no detectable off-target effects ... zebrafish Guide RNA expression plasmids for the EGFP Reporter Gene Guide RNA expression plasmids for ...