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Species: Homo sapiens
Genetic Insert: AAVS1 Homology Arms with CAGGS driven mEGFP
Vector Backbone Description: Vector Backbone:unknown; Vector Types:Mammalian Expression, CRISPR, Donor Template; Bacterial Resistance:Ampicillin
Comments: This plasmid has been used with locus-specific CRISPR/Cas9 to insert a CAGGS-mEGFP at the AAVS1 safe harbor locus in WTC human induced pluripotent stem cells by the Allen Institute for Cell Science. After insertion of the reporter using this donor template plasmid and CRISPR/Cas9 reagents, transfected cells may exhibit varying intensity levels of fluorescence, likely due to editing precision. To obtain cells of uniform intensity levels, see our protocol for fluorescence-assisted cell sorting and subcloning of transfected cells (https://www.allencell.org/instructional-videos-and-tutorials-for-cell-methods.html). Further, we recommend PCR-based assays for identifying precisely edited clones as previously described (https://www.molbiolcell.org/doi/abs/10.1091/mbc.e17-03-0209). For more information on the entire plasmid collection, please see https://www.addgene.org/allen-institute-cell-science/ .
Proper citation: RRID:Addgene_91565 Copy
Species: Homo sapiens
Genetic Insert: histone H2B-HaloTag
Vector Backbone Description: Backbone Marker:synthetic; Vector Backbone:pBR322; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:27484239
Proper citation: RRID:Addgene_91564 Copy
Vector Backbone Description: Backbone Marker:N/A; Backbone Size:4559; Vector Backbone:pSAM_Ec; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol and Kanamycin
Defining Citation: PMID:26060277
Comments: Mariner himar1C9 transposase driven by lac promoter on an R6K gamma suicide plasmid. The transposon allows identification of fitness factors by Illumina sequencing. This transposon will randomly insert a kanamycin resistance cassette into genomic TA sites. Chloramphenicol resistance is contained on the plasmid backbone.
For further background, see:
Goodman, Andrew L., Meng Wu, and Jeffrey I. Gordon. “Identifying Microbial Fitness Determinants by Insertion Sequencing (INSeq) Using Genome-Wide Transposon Mutant Libraries.” Nature Protocols 6.12 (2011): 1969–1980. PMC. Web. 11 Apr. 2017.
Wiles, Travis J. et al. “Combining Quantitative Genetic Footprinting and Trait Enrichment Analysis to Identify Fitness Determinants of a Bacterial Pathogen.” Ed. Danielle A. Garsin. PLoS Genetics 9.8 (2013): e1003716. PMC. Web. 11 Apr. 2017.
Proper citation: RRID:Addgene_91570 Copy
Species: Homo sapiens
Genetic Insert: CDH1 mutant promoter
Vector Backbone Description: Backbone Marker:Roel Nusse (Addgene plasmid # 24308); Backbone Size:9061; Vector Backbone:7TFP; Vector Types:Mammalian Expression, Lentiviral, Luciferase; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26941323
Proper citation: RRID:Addgene_91703 Copy
Species: Homo sapiens
Genetic Insert: CDH1 promoter
Vector Backbone Description: Backbone Marker:Roel Nusse (Addgene plasmid # 24308); Backbone Size:9061; Vector Backbone:7TFP; Vector Types:Mammalian Expression, Lentiviral, Luciferase; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26941323
Proper citation: RRID:Addgene_91704 Copy
Species: Homo sapiens
Genetic Insert: PHF2
Vector Backbone Description: Backbone Marker:David Root (Addgene plasmid # 41394); Backbone Size:9394; Vector Backbone:pLIX402; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26941323
Proper citation: RRID:Addgene_91701 Copy
Species: Homo sapiens
Genetic Insert: PHF2
Vector Backbone Description: Backbone Marker:David Root (Addgene plasmid # 41394); Backbone Size:9394; Vector Backbone:pLIX402; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26941323
Proper citation: RRID:Addgene_91700 Copy
Species: Homo sapiens
Genetic Insert: Human histone H2B
Vector Backbone Description: Backbone Marker:Trono lab; Backbone Size:7840; Vector Backbone:pRRL; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_91788 Copy
Species: Homo sapiens
Genetic Insert: Dectin1A
Vector Backbone Description: Backbone Marker:Trono lab; Backbone Size:8179; Vector Backbone:pRRL; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_91786 Copy
Species: Rattus norvegicus
Genetic Insert: GCaMP3-373
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:3955; Vector Backbone:pEGFP-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28162809
Proper citation: RRID:Addgene_91779 Copy
Species: Rattus norvegicus
Genetic Insert: GCaMP6-150
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:3955; Vector Backbone:pEGFP-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28162809
Proper citation: RRID:Addgene_91777 Copy
Species: Synthetic
Genetic Insert: mEGFP-P2A-paAIP2
Vector Backbone Description: Backbone Size:4650; Vector Backbone:pAAV-MCS; Vector Types:AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28318784
Proper citation: RRID:Addgene_91718 Copy
Species: Drosophila melanogaster
Genetic Insert: Laccase2
Vector Backbone Description: Vector Backbone:Hy_pMT; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29174924
Proper citation: RRID:Addgene_91799 Copy
Species: Synthetic
Genetic Insert: FKBP_F36V-2xHA-P2A-Puro
Vector Backbone Description: Backbone Marker:Takashi Yamamoto lab; Backbone Size:4579; Vector Backbone:pCRIS-PITChv2 (addgene #63672); Vector Types:Mammalian Expression, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29581585
Comments: See:
Sakuma, T., Nakade, S., Sakane, Y., Suzuki, K.-I. & Yamamoto, T. MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems. Nat Protoc 11, 118–133 (2016).
for the original backbone and general cloning protocol.
Proper citation: RRID:Addgene_91796 Copy
Vector Backbone Description: Backbone Marker:David Root Lab (addgene #41390); Backbone Size:9621; Vector Backbone:pLEX_305; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Chloramphenicol and Ampicillin
Defining Citation: PMID:29581585
Proper citation: RRID:Addgene_91797 Copy
Species: Synthetic
Genetic Insert: FKBP_F36V-2xHA-P2A-BSD
Vector Backbone Description: Backbone Marker:Takashi Yamamoto lab; Backbone Size:4579; Vector Backbone:pCRIS-PITChv2 (addgene #63672); Vector Types:Mammalian Expression, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29581585
Comments: See:
Sakuma, T., Nakade, S., Sakane, Y., Suzuki, K.-I. & Yamamoto, T. MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems. Nat Protoc 11, 118–133 (2016).
for the original backbone and general cloning protocol.
Proper citation: RRID:Addgene_91795 Copy
Species: Synthetic
Genetic Insert: Puro-P2A-2xHA-FKBP_F36V
Vector Backbone Description: Backbone Marker:Takashi Yamamoto lab; Backbone Size:4579; Vector Backbone:pCRIS-PITChv2 (addgene #63672); Vector Types:Mammalian Expression, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29581585
Comments: See:
Sakuma, T., Nakade, S., Sakane, Y., Suzuki, K.-I. & Yamamoto, T. MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems. Nat Protoc 11, 118–133 (2016).
for the original backbone and general cloning protocol.
Proper citation: RRID:Addgene_91793 Copy
Species: Streptococcus pyogenes
Genetic Insert: SpCas9
Vector Backbone Description: Backbone Size:17766; Vector Backbone:pZD_OsU3gYSA_HolgerCas9_NPTII; Vector Types:Plant Expression; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:28346401
Proper citation: RRID:Addgene_91695 Copy
Species: Discosoma sp. (mushroom coral)
Genetic Insert: monomeric Cherry
Vector Backbone Description: Backbone Marker:Life Technologies; Backbone Size:2900; Vector Backbone:modified pRSETB; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15558047
Comments: These plasmids are used in University Molecular Biology Laboratories and by High School students to purify fluorescent proteins and create fluorescent protein plate artwork.
Addgene next-generation sequencing identified a deletion of Glutimate and Glycine residues (amino acids 73 and 74 relative to Genbank ID: 6MZ3_A). The depositing laboratory has observed expression of the chromophore in E. coli.
Proper citation: RRID:Addgene_91769 Copy
Species: Discosoma sp. (mushroom coral)
Genetic Insert: tandem dimer Tomato
Vector Backbone Description: Backbone Marker:Life Technologies; Backbone Size:2900; Vector Backbone:modified pRSETB; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15558047
Comments: These plasmids are used in University Molecular Biology Laboratories and by High School students to purify fluorescent proteins and create fluorescent protein plate artwork.
Proper citation: RRID:Addgene_91767 Copy
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