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doxycycline induced promoter (Addgene inc)

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Addgene inc doxycycline induced promoter
Doxycycline Induced Promoter, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 372 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/doxycycline+inducible+promoter/pInducer20+(Plasmid+%2344012)/bio_rxiv__64898__2026__01__13__699073-237-13-20
Average 96 stars, based on 372 article reviews

doxycycline induced promoter - by Bioz Stars, 2026-09

96/100 stars

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Expressing:

Article Title: Cell-type-specific adaptations to mitochondrial stress underly the neurological presentations of MTRFR mutations
Article Snippet: Reprogramming vectors pCXLE-hOct3/4 shp53-F (Addgene #27077), pCXLE-hSK (Addgene #27078), pCXLE-hUL (Addgene #27080) and pCXLE-eGPP (Addgene #27082) were gifts from Shinya Yamanaka. .. A lentivirus expressing Ngn2 (pTet-O-Ngn2-puro; Addgene: #52047) and a doxycycline-inducible promoter (FUdeltaGW-rtTA; Addgene: #19780) was generated using the pPACKH1 HIV Lentivector Packaging Kit (System Bio) following manufacturer’s instructions using HEK293 cells. .. The virus was precipitated with the PEG-it Virus Precipitation Solution (System Biosciences).

Article Title: Complete human day 14 post-implantation embryo models from naive ES cells
Article Snippet: .. To generate Tet-ON inducible lines, we used a PiggyBac plasmid expressing a cDNA insert and transposase vector of choice under the control of a doxycycline-inducible promoter (a gift from V. Busskamp, Addgene, plasmid 104454). ..

Article Title: mRNA interactions with disordered regions control protein activity
Article Snippet: .. Doxycycline inducible Cas9 (iCas9) HeLa cells were generated by infecting cells with lentivirus containing a Cas9-P2A-GFP expression cassette under a doxycycline inducible promoter as described previously (Addgene plasmid #85400) . ..

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Article Title: HP1 loses its chromatin clustering and phase separation function across evolution
Article Snippet: .. HP1 variants are expressed from a doxycycline inducible promoter (plasmid pSBtet-BP, referenced in the Methods section along with its Addgene number and the primary reference). ..

Article Title: Whole-genome screens reveal regulators of differentiation state and context-dependent migration in human neutrophils
Article Snippet: This construct was genomically integrated by lentiviral transduction following the same method described for the construct of the main text. c dCas9-BFP-KRAB (Addgene plasmid #46911). .. This construct was genomically integrated by lentiviral transduction following the same method described for the construct of the main text. d dCas9-KRAB-P2A-mCherry driven by a doxycycline Inducible promoter (Addgene plasmid #73497). ..

Article Title: A CRISPR/Cas9-Based System for Reprogramming Cell Lineage Specification
Article Snippet: .. The lentiviral backbone along with the doxycycline inducible promoter was derived from Tet-O-FUW-Myt1l (Addgene plasmid 27152) (Vierbuchen et al., 2010). ..

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( A ) Heatmap showing the top 50 differentially regulated genes from RNA-Seq data. Unsupervised hierarchical clustering by sample and gene were performed. The data are based on samples from the F-MPN ( n = 7) and control ( n = 4) MSC groups. In the top 50 genes, boxed genes are associated with either fibrosis or osteoblast differentiation pathways. ( B ) Gene accessibility tracks of HOXB genes (ATAC-Seq) show a statistically significant increased accessibility in F-MPNs (example of the HOXB-AS3-HOXB7 region is quantified in the right panel). P value *< 0.02 calculated using unpaired t test. ( C ) qRT-PCR validation of the ATAC-Seq and RNA-Seq findings in control ( n = 13) and F-MPN ( n = 10) samples. Several HOXB genes are upregulated in F-MPN MSCs. P values (*<0.05, **<0.01) were calculated using unpaired t test. ( D ) In vitro coculture of normal human mesenchymal cells from the HS-5 cell line with either cells or conditioned media from human hematopoietic cell line UT-7 bearing either the wild-type JAK2 gene (WT) or the mutated JAK2 V617F gene (VF) or with TGF-β (10 ng/mL). ( E ) After 7 days’ coculture, an increased expression of HOXB7 and ACTA2 genes is noted in the MSCs after exposure to TGF-β or JAK2 -mutated cells (VF). P values (*<0.05) were calculated using unpaired t test.

Journal: JCI Insight

Article Title: Comprehensive analysis of mesenchymal cells reveals a dysregulated TGF- β /WNT/HOXB7 axis in patients with myelofibrosis

doi: 10.1172/jci.insight.173665

Figure Lengend Snippet: ( A ) Heatmap showing the top 50 differentially regulated genes from RNA-Seq data. Unsupervised hierarchical clustering by sample and gene were performed. The data are based on samples from the F-MPN ( n = 7) and control ( n = 4) MSC groups. In the top 50 genes, boxed genes are associated with either fibrosis or osteoblast differentiation pathways. ( B ) Gene accessibility tracks of HOXB genes (ATAC-Seq) show a statistically significant increased accessibility in F-MPNs (example of the HOXB-AS3-HOXB7 region is quantified in the right panel). P value *< 0.02 calculated using unpaired t test. ( C ) qRT-PCR validation of the ATAC-Seq and RNA-Seq findings in control ( n = 13) and F-MPN ( n = 10) samples. Several HOXB genes are upregulated in F-MPN MSCs. P values (*<0.05, **<0.01) were calculated using unpaired t test. ( D ) In vitro coculture of normal human mesenchymal cells from the HS-5 cell line with either cells or conditioned media from human hematopoietic cell line UT-7 bearing either the wild-type JAK2 gene (WT) or the mutated JAK2 V617F gene (VF) or with TGF-β (10 ng/mL). ( E ) After 7 days’ coculture, an increased expression of HOXB7 and ACTA2 genes is noted in the MSCs after exposure to TGF-β or JAK2 -mutated cells (VF). P values (*<0.05) were calculated using unpaired t test.

Article Snippet: Doxycycline-inducible HOXB7 shRNA was generated using Tet-pLKO-puro plasmid (Addgene plasmid 21915; https://www.addgene.org/21915/ ).

Techniques: RNA Sequencing, Control, Quantitative RT-PCR, Biomarker Discovery, In Vitro, Expressing

( A ) Knockdown of HOXB7 by shRNA in the HS-5 mesenchymal cell line. The reduced expression of HOXB7 is validated in an shRNA-transfected cell line by qRT-PCR. ( B ) Knockdown of HOXB7 by shRNA in the HS-5 mesenchymal cell line. Western blot showing absence of HOXB7 in 2 shRNA-transfected cell lines results in downregulation of the downstream target α-SMA (protein encoded by the ACTA2 gene). ( C ) Gene accessibility tracks of 2 osteoblast-associated genes, ACTA2 and TAGLN , by ATAC-Seq. HS-5 cells with reduced expression of HOXB7 (HOXB7-KD3) show reduced chromatin accessibility of the ACTA2 and TAGLN genes even after incubation with TGF-β (10 ng/mL for 3 days). ( D ) qRT-PCR validation of the ATAC-Seq findings showing significantly reduced expression of the ACTA2 and TAGLN genes in the HOXB7 -KD3 cells in the presence of TGF-β. P values *<0.05, **<0.02 calculated using unpaired t test. ( E ) qRT-PCR results showing the downregulation of HOXB7 and TAGLN even in the presence of TGF-β upon doxycycline (Dox) addition, and the same is restored when the Dox is withdrawn from the same cells after 3 days ( n = 3). ( F ) Osteoblast differentiation induction assay of HS-5 cells after transfection with either scramble (SCR) or HOXB7 shRNA ( HOXB7 KD3) alone or after treatment with TGF-β for 21 days. (Left panel: microscopy images after osteoblast lineage staining; original magnification is ×10; right panel: arbitrary quantification of osteoblast differentiation.) Absence of HOXB7 reduces osteoblast differentiation even in the presence of TGF-β. P values were calculated using 1-way ANOVA test.

Journal: JCI Insight

Article Title: Comprehensive analysis of mesenchymal cells reveals a dysregulated TGF- β /WNT/HOXB7 axis in patients with myelofibrosis

doi: 10.1172/jci.insight.173665

Figure Lengend Snippet: ( A ) Knockdown of HOXB7 by shRNA in the HS-5 mesenchymal cell line. The reduced expression of HOXB7 is validated in an shRNA-transfected cell line by qRT-PCR. ( B ) Knockdown of HOXB7 by shRNA in the HS-5 mesenchymal cell line. Western blot showing absence of HOXB7 in 2 shRNA-transfected cell lines results in downregulation of the downstream target α-SMA (protein encoded by the ACTA2 gene). ( C ) Gene accessibility tracks of 2 osteoblast-associated genes, ACTA2 and TAGLN , by ATAC-Seq. HS-5 cells with reduced expression of HOXB7 (HOXB7-KD3) show reduced chromatin accessibility of the ACTA2 and TAGLN genes even after incubation with TGF-β (10 ng/mL for 3 days). ( D ) qRT-PCR validation of the ATAC-Seq findings showing significantly reduced expression of the ACTA2 and TAGLN genes in the HOXB7 -KD3 cells in the presence of TGF-β. P values *<0.05, **<0.02 calculated using unpaired t test. ( E ) qRT-PCR results showing the downregulation of HOXB7 and TAGLN even in the presence of TGF-β upon doxycycline (Dox) addition, and the same is restored when the Dox is withdrawn from the same cells after 3 days ( n = 3). ( F ) Osteoblast differentiation induction assay of HS-5 cells after transfection with either scramble (SCR) or HOXB7 shRNA ( HOXB7 KD3) alone or after treatment with TGF-β for 21 days. (Left panel: microscopy images after osteoblast lineage staining; original magnification is ×10; right panel: arbitrary quantification of osteoblast differentiation.) Absence of HOXB7 reduces osteoblast differentiation even in the presence of TGF-β. P values were calculated using 1-way ANOVA test.

Article Snippet: Doxycycline-inducible HOXB7 shRNA was generated using Tet-pLKO-puro plasmid (Addgene plasmid 21915; https://www.addgene.org/21915/ ).

Techniques: Knockdown, shRNA, Expressing, Transfection, Quantitative RT-PCR, Western Blot, Incubation, Biomarker Discovery, Microscopy, Staining

Journal: JCI Insight

Article Title: Comprehensive analysis of mesenchymal cells reveals a dysregulated TGF- β /WNT/HOXB7 axis in patients with myelofibrosis

doi: 10.1172/jci.insight.173665

Figure Lengend Snippet: ( A ) Treatment of normal human mesenchymal cells (HS-5) with TGF-β (2 and 10 ng/mL) stabilizes β-catenin levels (immunoblot). ( B ) Treatment of normal human mesenchymal cells (HS-5) with TGF-β (10 ng/mL) induces translocation of β-catenin to the nucleus (red arrows). Microscopy images after staining with mAb-β-catenin (green) and DAPI (blue nucleus) (representative of 3 experiments). Original magnification, ×100. ( C ) ChIP assay of the HOXB7 promoter. Incubation with TGF-β (10 ng/mL) drives increased chromatin accessibility (reduced H3K9me3 marks) and binding of β-catenin ( n = 3) to the HOXB7 promoter. P value *<0.05, unpaired t test. ( D ) Knockdown of β-catenin in HS-5 cell line by a β-catenin shRNA (β-Cat KD), validated by Western blot (right), induces reduced expression of HOXB7 and its downstream target α-SMA (protein encoded by ACTA2 ) in the absence or presence of TGF-β. ( E ) Osteoblast and adipocyte differentiation induction assay of normal mesenchymal HS-5 cells after transfection with scramble (SCR) or β-catenin shRNA (β-Cat KD). Microscopy images after respective lineage staining show that absence of β-catenin decreases osteoblast differentiation and increases adipocyte differentiation. Original magnification, ×10. ( F ) Pretreatment of HS-5 cells with WNT inhibitor (cardamonin 20 μM) followed by treatment with TGF-β reduces the expression of HOXB7 and α-SMA along with other WNT pathway genes such as c-MYC and Cyclin-D1 ( n = 3). ( G ) Treatment of HS-5 cells with TGF-β and WNT inhibitor reduces the expression of ACTA2 and TAGLN genes validated by qRT-PCR ( n = 3). P value **<0.02, unpaired t test. ( H ) Osteodifferentiation induction of HS-5 cells shows a decreased osteoblast differentiation profile upon treatment by cardamonin ( n = 3). Original magnification, ×10. P value *<0.05, unpaired t test.

Article Snippet: Doxycycline-inducible HOXB7 shRNA was generated using Tet-pLKO-puro plasmid (Addgene plasmid 21915; https://www.addgene.org/21915/ ).

Techniques: Western Blot, Translocation Assay, Microscopy, Staining, Incubation, Binding Assay, Knockdown, shRNA, Expressing, Transfection, Quantitative RT-PCR

Biochemically-gated imaging of transcriptional activity. (a) Gated reporters leverage genetic AND logic to generate an output only when transcriptional activity ( T x ) is accompanied by an externally added drug (in this case, shield-1). (b) Doxycycline-dependent change in diffusivities of CHO cells expressing conventional DD-free Aqp1 under the control of a doxycycline-inducible minimal CMV promoter. (c) Doxycycline-driven changes in diffusivities of CHO cells expressing Aqp1-FKBP12-DD under the control of a doxycycline-inducible minimal CMV promoter following incubation with shield-1, doxycycline, and both ligands. (d) Representative difference image showing shield-1 gated imaging of transcriptional activity. The difference image was obtained through voxel-wise subtraction of diffusion-weighted datasets (effective b -value ∼1.6 ms μm −2 ) acquired in the presence of shield-1, doxycycline, or both from a dataset acquired in their absence. The resulting image was subsequently denoised using a median filter and presented as a pseudo-colored “hotspot.” Error bars represent the standard deviation ( n = 3–6). *** denotes P < 0.001. P -values were computed based on 2-sided, unpaired t -test (2b) or using one-way ANOVA followed by Tukey's HSD test (2c).

Journal: Chemical Science

Article Title: Destabilized reporters for background-subtracted, chemically-gated, and multiplexed deep-tissue imaging †

doi: 10.1039/d4sc00377b

Figure Lengend Snippet: Biochemically-gated imaging of transcriptional activity. (a) Gated reporters leverage genetic AND logic to generate an output only when transcriptional activity ( T x ) is accompanied by an externally added drug (in this case, shield-1). (b) Doxycycline-dependent change in diffusivities of CHO cells expressing conventional DD-free Aqp1 under the control of a doxycycline-inducible minimal CMV promoter. (c) Doxycycline-driven changes in diffusivities of CHO cells expressing Aqp1-FKBP12-DD under the control of a doxycycline-inducible minimal CMV promoter following incubation with shield-1, doxycycline, and both ligands. (d) Representative difference image showing shield-1 gated imaging of transcriptional activity. The difference image was obtained through voxel-wise subtraction of diffusion-weighted datasets (effective b -value ∼1.6 ms μm −2 ) acquired in the presence of shield-1, doxycycline, or both from a dataset acquired in their absence. The resulting image was subsequently denoised using a median filter and presented as a pseudo-colored “hotspot.” Error bars represent the standard deviation ( n = 3–6). *** denotes P < 0.001. P -values were computed based on 2-sided, unpaired t -test (2b) or using one-way ANOVA followed by Tukey's HSD test (2c).

Article Snippet: Plasmids harboring the various degron sequences – DHFR-DD (Addgene 29326), ER-DD (Addgene 37261), miniIAA7 (Addgene 129721), and FKBP12 (Addgene 17416) were amplified using Q5 High-Fidelity 2× Master Mix and cloned by Gibson assembly in a lentiviral transfer vector at the C or N -terminus of the aquaporin-1 reporter (Aqp1) under the control of either a constitutive promoter, EF1α (Addgene 60058) or a doxycycline-inducible minimal CMV promoter (Addgene 26431).

Techniques: Imaging, Activity Assay, Expressing, Control, Incubation, Diffusion-based Assay, Standard Deviation

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