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http://www.addgene.org/216385

Species: Homo sapiens
Genetic Insert: Norrin
Vector Backbone Description: Backbone Marker:BD Biosciences; Vector Backbone:pAcGP67a; Vector Types:Insect Expression, Baculovirus; Bacterial Resistance:Ampicillin
Comments: Please visit https://doi.org/10.1101/2024.02.03.578714 for bioRxiv preprint.

Proper citation: RRID:Addgene_216385 Copy   

•   Source: Addgene

http://www.addgene.org/216384

Species: Homo sapiens
Genetic Insert: Norrin
Vector Backbone Description: Backbone Marker:BD Biosciences; Vector Backbone:pAcGP67a; Vector Types:Insect Expression, Baculovirus; Bacterial Resistance:Ampicillin
Comments: Please visit https://doi.org/10.1101/2024.02.03.578714 for bioRxiv preprint.

Proper citation: RRID:Addgene_216384 Copy   

•   Source: Addgene

http://www.addgene.org/217640

Species: Homo sapiens
Genetic Insert: WT1 +KTS isoform
Vector Backbone Description: Vector Backbone:pENTR; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:33882316

Proper citation: RRID:Addgene_217640 Copy   

•   Source: Addgene

http://www.addgene.org/216791

Species: Homo sapiens
Genetic Insert: MKK6
Vector Backbone Description: Vector Backbone:pBABE puro; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:39536108
Comments: This plasmid was created by PCR cloning pGEX-4T-1 3xHA-MKK6-EE (Addgene #47579) using BglII (5') and SalI (3') and inserting into pBABE puro that had been digested by BamHI (5') and SalI (3')

Proper citation: RRID:Addgene_216791 Copy   

•   Source: Addgene

http://www.addgene.org/217641

Species: Homo sapiens
Genetic Insert: WT1 -KTS isoform
Vector Backbone Description: Vector Backbone:pENTR; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:33882316

Proper citation: RRID:Addgene_217641 Copy   

•   Source: Addgene

http://www.addgene.org/217645

Species: Homo sapiens
Genetic Insert: WT1
Vector Backbone Description: Backbone Marker:Addgene #41393; Vector Backbone:pCW57.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33882316

Proper citation: RRID:Addgene_217645 Copy   

•   Source: Addgene

http://www.addgene.org/217521

Species: Homo sapiens
Genetic Insert: tdTomato-tDeg fluorogenic protein
Vector Backbone Description: Backbone Size:1581; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:38296979

Proper citation: RRID:Addgene_217521 Copy   

•   Source: Addgene

http://www.addgene.org/217642

Species: Homo sapiens
Genetic Insert: WT1 -KTS isoform
Vector Backbone Description: Vector Backbone:pENTR; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:33882316

Proper citation: RRID:Addgene_217642 Copy   

•   Source: Addgene

http://www.addgene.org/216570

Species: Homo sapiens
Genetic Insert: TCF4
Vector Backbone Description: Backbone Marker:GenScript; Backbone Size:2800; Vector Backbone:pCR8-GW-TOPO; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Comments: insert contains a Kozak sequence that encodes a linker for cloning into N-terminal tagging pDEST vectors

Proper citation: RRID:Addgene_216570 Copy   

•   Source: Addgene

http://www.addgene.org/217659

Species: Homo sapiens
Genetic Insert: sgRNA2 targeting the human AAVS1 safe harbour locus
Vector Backbone Description: Backbone Size:9643; Vector Backbone:pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem-Bpil-P2A-mCherry; Vector Types:Mammalian Expression, Bacterial Expression, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:37357575

Proper citation: RRID:Addgene_217659 Copy   

•   Source: Addgene

http://www.addgene.org/217418

Species: Homo sapiens
Genetic Insert: Luciferase, GFP
Vector Backbone Description: Backbone Marker:Boris Fehse (Addgene #27358); Vector Backbone:LeGO-iG; Vector Types:Mammalian Expression, Lentiviral, Luciferase; Bacterial Resistance:Ampicillin
Comments: This is an inducible lentiviral luciferase expression vector that has 6 RAS/MAPK-inducible AP-1 sites driving transcription from the human DUSP5 promoter, which is highly active in most acute myeloid leukaemia samples but not in normal myeloid stem cells (Assi et al, 2019). This is a CG island promoter containing Sp1, SRF, ETS and NF-kB sites, and is likely to be activated by signalling downstream of FLT3-ITD mutations (Assi et al, 2019). This vector also contains a myeloid cell chromatin priming element from the human IL3 gene that can potentially diminish chromatin silencing effects by maintaining open chromatin. LEGO-pDHS-AP1x6-DUSP5 was constructed from LEGO-pDHS-AP1x6-GM55 by replacing the minimal human CSF2 promoter with the full-length human DUSP5 promoter. The pDHS was added by inserting into the XbaI site a 274 bp PCR-generated fragment of a DNA segment located 10 kb upstream of the human IL3 gene that maintains open chromatin in myeloid cells. LEGO-AP1x6-GM55 was previously constructed from LeGO-iG (Weber et al, 2008) (Addgene #27358) by replacing a NotI/XbaI fragment containing the U6 and SFFV promoters with a fragment from the plasmid pAP1-3 (Addgene #71258) that has 3 copies of the MMP3 gene AP-1 site in front of human CSF2 -55 to +28 minimal promoter driving expression of the Luc+ firefly luciferase gene originating from the plasmid pGL3 (Promega), and then duplicating the array of 3 AP-1 sites. The luciferase gene is linked to an IRES that in principle supports co-expression of luciferase and EGFP. The insert has 2 copies of the SV40 transcription termination/polyadenylation signal upstream of the multiple cloning site, which blocks background read-through transcription originating in the vector backbone, which itself can be substantial (Bert et al, 2000). The inserted luciferase gene and SV40 polyA site sequences originate from the plasmid pXPG (Bert et al, 2000) (Addgene #71248). The priming element (pDHS) encompasses RUNX1 and ETS1 binding sites and resembles sites that prime genes for rapid activation in T cells (Bevington et al, 2016). References: Assi et al. Nat Genet. 2019; 51:151-162. PMID 30420649. Bert et al. Plasmid. 2000 Sep;44(2):173-82. PMID 10964627. Weber et al. Mol Ther. 2008 Apr. 16(4):698-706. PMID 18362927. Bevington et al. Bioessays 2016 Dec 27. doi: 10.1002/bies.201600184. PMID 28026028.

Proper citation: RRID:Addgene_217418 Copy   

•   Source: Addgene

http://www.addgene.org/216724

Species: Homo sapiens
Genetic Insert: KIF17
Vector Backbone Description: Vector Backbone:pEGFP-N3; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:37640938

Proper citation: RRID:Addgene_216724 Copy   

•   Source: Addgene

http://www.addgene.org/217416

Species: Homo sapiens
Genetic Insert: Luciferase, GFP
Vector Backbone Description: Backbone Marker:Boris Fehse (Addgene #27358); Vector Backbone:LeGO-iG; Vector Types:Mammalian Expression, Lentiviral, Luciferase; Bacterial Resistance:Ampicillin
Comments: This is an inducible lentiviral luciferase expression vector that has 6 RAS/MAPK-inducible AP-1 sites driving transcription from the minimal CSF2 promoter, plus a chromatin priming element from the IL3 gene that can potentially diminish chromatin silencing effects. LEGO-pDHS-AP1x6-GM55 was constructed from LEGO-AP1x6-GM55 by inserting into the XbaI site a 274 bp PCR-generated fragment of a DNA segment located 10 kb upstream of the human IL3 gene that maintains open chromatin at a constitutive DNaseI Hypersensitive Site in myeloid cells. LEGO-AP1x6-GM55 was constructed from LeGO-iG (Weber et al, 2008) (Addgene #27358) by replacing a NotI/XbaI fragment containing the U6 and SFFV promoters with a fragment from the plasmid pAP1-3 (Addgene #71258) that has 3 copies of the MMP3 gene AP-1 site in front of human CSF2 -55 to +28 minimal promoter driving expression of the Luc+ firefly luciferase gene originating from the plasmid pGL3 (Promega), and then duplicating the array of 3 AP-1 sites. The luciferase gene is linked to an IRES that in principle supports co-expression of luciferase and EGFP. The insert has 2 copies of the SV40 transcription termination/polyadenylation signal upstream of the multiple cloning site, which blocks background read-through transcription originating in the vector backbone, which itself can be substantial (Bert et al, 2000). The inserted luciferase gene and SV40 polyA site sequences originate from the plasmid pXPG (Bert et al, 2000) (Addgene #71248). The priming element (pDHS) encompasses RUNX1 and ETS1 binding sites and resembles sites that prime genes for rapid activation in T cells (Bevington et al, 2016). References: Bert et al. Plasmid. 2000 Sep;44(2):173-82. PMID 10964627. Weber et al. Mol Ther. 2008 Apr. 16(4):698-706. PMID 18362927. Bevington et al. Bioessays 2016 Dec 27. doi: 10.1002/bies.201600184. PMID 28026028.

Proper citation: RRID:Addgene_217416 Copy   

•   Source: Addgene

http://www.addgene.org/217419

Species: Homo sapiens
Genetic Insert: Luciferase, GFP
Vector Backbone Description: Backbone Marker:Boris Fehse (Addgene #27358); Vector Backbone:LeGO-iG; Vector Types:Mammalian Expression, Lentiviral, Luciferase; Bacterial Resistance:Ampicillin
Comments: This expression vector contains a synthetic enhancer that was designed as an array of transcription factor binding sites predicted to be highly active in acute myeloid leukaemia cells carrying activating mutations in the receptor FLT3 (Assi et al., 2019). This putative enhancer was generated as an oligonucleotide duplex containing binding sites for EGR1, ETS, SRF, CREB, NF1, NF-kB and WT1, inserted into LEGO-pDHS-AP1x6-DUSP5. The closely linked ETS, SRF, and CREB sites are derived from a Serum Response Element in the FOS promoter which is induced by MAPK. LEGO-pDHS-AP1x6-DUSP5 is an inducible lentiviral luciferase expression vector that can has 6 RAS/MAPK-inducible AP-1 sites driving transcription from the human DUSP5 promoter, which is highly active in most acute myeloid leukaemia samples but not in normal myeloid stem cells (Assi et al, 2019). This is a CG island promoter containing Sp1, SRF, ETS and NF-kB sites, and is likely to be activated by signalling downstream of FLT3-ITD mutations (Assi et al, 2019). This vector also contains a myeloid cell chromatin priming element from the human IL3 gene that can potentially diminish chromatin silencing effects by maintaining open chromatin. LEGO-pDHS-AP1x6-DUSP5 was constructed from LEGO-pDHS-AP1x6-GM55 by replacing the minimal human CSF2 promoter with the full-length human DUSP5 promoter. The pDHS was added by inserting into the XbaI site a 274 bp PCR-generated fragment of a DNA segment located 10 kb upstream of the human IL3 gene that maintains open chromatin in myeloid cells. LEGO-AP1x6-GM55 was previously constructed from LeGO-iG (Weber et al, 2008) (Addgene #27358) by replacing a NotI/XbaI fragment containing the U6 and SFFV promoters with a fragment from the plasmid pAP1-3 (Addgene #71258) that has 3 copies of the MMP3 gene AP-1 site in front of human CSF2 -55 to +28 minimal promoter driving expression of the Luc+ firefly luciferase gene originating from the plasmid pGL3 (Promega), and then duplicating the array of 3 AP-1 sites. The luciferase gene is linked to an IRES that in principle supports co-expression of luciferase and EGFP. The insert has 2 copies of the SV40 transcription termination/polyadenylation signal upstream of the multiple cloning site, which blocks background read-through transcription originating in the vector backbone, which itself can be substantial (Bert et al, 2000). The inserted luciferase gene and SV40 polyA site sequences originate from the plasmid pXPG (Bert et al, 2000) (Addgene #71248). The priming element (pDHS) encompasses RUNX1 and ETS1 binding sites and resembles sites that prime genes for rapid activation in T cells (Bevington et al, 2016)). References: Assi et al. Nat Genet. 2019; 51:151-162. PMID 30420649. Bert et al. Plasmid. 2000 Sep;44(2):173-82. PMID 10964627. Weber et al. Mol Ther. 2008 Apr. 16(4):698-706. PMID 18362927. Bevington et al. Bioessays 2016 Dec 27. doi: 10.1002/bies.201600184. PMID 28026028.

Proper citation: RRID:Addgene_217419 Copy   

•   Source: Addgene

http://www.addgene.org/217013

Species: Homo sapiens
Genetic Insert: CDK7(W132R, S164D, T170E)
Vector Backbone Description: Vector Backbone:pFastBac dual; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin and Gentamicin
Defining Citation: PMID:38870939

Proper citation: RRID:Addgene_217013 Copy   

•   Source: Addgene

http://www.addgene.org/217670

Species: Homo sapiens
Genetic Insert: CRYAB
Vector Backbone Description: Backbone Marker:LifeSensors; Backbone Size:5761; Vector Backbone:PE-SUMO; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:39368693
Comments: Expressed in a soluble state, except when grown in minimal media to produce 15N labeled protein. Can be purified using N-terminal 6xHis tag and SUMO tag can then be efficiently removed by ULP-1 cleavage yielding the N-terminal methionine.

Proper citation: RRID:Addgene_217670 Copy   

•   Source: Addgene

http://www.addgene.org/217666

Species: Homo sapiens
Genetic Insert: WapI
Vector Backbone Description: Backbone Size:3913; Vector Backbone:pCR2.1; Vector Types:Mammalian Expression, Bacterial Expression; Bacterial Resistance:Ampicillin and Kanamycin
Defining Citation: PMID:37357575

Proper citation: RRID:Addgene_217666 Copy   

•   Source: Addgene

http://www.addgene.org/216624

Species: Homo sapiens
Genetic Insert: STXBP1
Vector Backbone Description: Backbone Marker:GenScript; Backbone Size:2800; Vector Backbone:pCR8-GW-TOPO; Vector Types:Gateway Cloning; Bacterial Resistance:Spectinomycin
Comments: insert contains a Kozak sequence that encodes a linker for cloning into N-terminal tagging pDEST vectors

Proper citation: RRID:Addgene_216624 Copy   

•   Source: Addgene

http://www.addgene.org/217398

Species: Homo sapiens
Genetic Insert: hM4D(Gi)-mCherry
Vector Backbone Description: Backbone Size:4923; Vector Backbone:pAAV; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin

Proper citation: RRID:Addgene_217398 Copy   

•   Source: Addgene

http://www.addgene.org/217673

Species: Homo sapiens
Genetic Insert: CRYGS
Vector Backbone Description: Backbone Size:5368; Vector Backbone:pET28B+; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: This a N-terminally truncated gamma S crystallin missing its N-terminal 15 amino acid residues and containing a Q16C mutation at its N-terminus to act as a site to perform native chemical ligation. It also has a N-terminal SUMO tag with a 6X His tag with the amino acid sequence: GHHHHHHGSLQDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRRLMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQ IGG It will express in a soluble state with the SUMO tag, but will precipitate once the SUMO tag is removed during ulp-1 cleavage. It has a propensity to form non-disulfide linked multimers once the SUMO tag is removed, so the ligation reaction must be performed immediately after ULP-1 treatment.

Proper citation: RRID:Addgene_217673 Copy   

•   Source: Addgene