Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

A chemical-induced reversible gene knockout system for studying gene function in situ [RNA-seq]

ABSTRACT: We developed a conditional conversion allele system termed CoCo. Gene knockout is achieved through Cre-mediated transcription termination, whereas gene restoration is achieved by Flp-mediated deletion of termination signal. To tightly control the temporal expression of the recombinases, Tet-On inducible system is used to drive the expression of Cre recombinase and tamoxifen-inducible FlpO-ER fusion and therefore a dual drug-inducible conditional conversion system (DDCoCo) is developed. This system provides an efficient, rapid and flexible gene switch for studying gene function.

ORGANISM(S): Homo sapiens

PROVIDER: GSE188384 | GEO | 2021/11/13

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

A chemical-induced reversible gene knockout system for studying gene function in situ [AMP-seq]

Project description:We developed a conditional conversion allele system termed CoCo. Gene knockout is achieved through Cre-mediated transcription termination, whereas gene restoration is achieved by Flp-mediated deletion of termination signal. To tightly control the temporal expression of the recombinases, Tet-On inducible system is used to drive the expression of Cre recombinase and tamoxifen-inducible FlpO-ER fusion and therefore a dual drug-inducible conditional conversion system (DDCoCo) is developed. This system provides an efficient, rapid and flexible gene switch for studying gene function.

2021-11-13 | GSE188594 | GEO

Proteome analysis of GCN1 CKO mouse ileum

Project description:Tamoxifen-inducible conditional knockout (CKO) mice were generated to explore the function of Gcn1 in adult mice using the Cre/loxP system. To analyze the function of GCN1 in the intestinal epithelium, we compared the whole cell proteome of ileum harvested from GCN1 CKO mice with that of wild-type mice.

2024-03-31 | PXD030518 | JPOST Repository

Proteome analysis of GCN1 CKO mouse duodenum

Project description:Tamoxifen-inducible conditional knockout (CKO) mice were generated to explore the function of Gcn1 in adult mice using the Cre/loxP system. To analyze the function of GCN1 in the intestinal epithelium, we compared the whole cell proteome of duodenum harvested from GCN1 CKO mice with that of wild-type mice.

2024-03-31 | PXD030517 | JPOST Repository

Proteome analysis of GCN1 CKO mouse liver

Project description:Tamoxifen-inducible conditional knockout (CKO) mice were generated to explore the function of Gcn1 in adult mice using the Cre/loxP system. To analyze the function of GCN1 in the liver, we compared the whole cell proteome of livers harvested from GCN1 CKO mice with that of wild-type mice.

2023-04-01 | PXD030514 | JPOST Repository

Proteome analysis of GCN1 CKO mouse jejunum

Project description:Tamoxifen-inducible conditional knockout (CKO) mice were generated to explore the function of Gcn1 in adult mice using the Cre/loxP system. To analyze the function of GCN1 in the intestinal epithelium, we compared the whole cell proteome of jejunum harvested from GCN1 CKO mice with that of wild-type mice.

2022-08-29 | PXD030519 | JPOST Repository

Proteome analysis of GCN1 CKO mouse inguinal white adipose tissue (iWAT)

Project description:Tamoxifen-inducible conditional knockout (CKO) mice were generated to explore the function of Gcn1 in adult mice using the Cre/loxP system. To analyze the function of GCN1 in the inguinal white adipose tissue (iWAT), we compared the whole cell proteome of eWAT harvested from GCN1 CKO mice with that of wild-type mice.

2024-03-31 | PXD030516 | JPOST Repository

Engineering spacer specificity of the Cre/loxP system

Project description:Translational research on the Cre/loxP recombination system focuses on enhancing its specificity by modifying Cre/DNA interactions. Despite extensive efforts, the exact mechanisms governing how Cre distinguishes between substrates remains elusive. Cre recognizes 13 bp inverted repeats, initiating recombination in the 8 bp spacer region. While literature suggests that efficient recombination proceeds between lox sites with non-loxP spacer sequences when both lox sites have matching spacers, experimental validation for this assumption is lacking. To fill this gap, we investigated target site variations of identical pairs of the loxP 8 bp spacer region, screening 6,000 unique loxP-like sequences. Approximately 84% of these sites exhibited efficient recombination, affirming the flexibility of spacer sequences for catalysis. However, certain spacers negatively impacted recombination, emphasizing sequence dependence. Directed evolution of Cre on inefficiently recombined spacers not only yielded recombinases with enhanced activity but also mutants with reprogrammed selective activity. Mutations altering spacer specificity were identified, and molecular modelling and dynamics simulations elucidated the mechanism behind this specificity switch. These findings highlight the potential to fine-tune site-specific recombinases for spacer sequence specificity, offering a novel concept to enhance the applied properties of designer-recombinases for genome engineering applications.

2024-07-25 | GSE254392 | GEO

Cre-recombinase expression cooperates with FLT3ITD/ITD to induce acute myeloid leukemia [RNASeq_SclCre_Spleen]

Project description:Animal models offer a valuable tool to recapitulate genetically defined subtypes of AML, and to assess the potential of compound mutations and clonal evolution during disease progression. This is of utmost importance for difficult to treat leukemias such as FLT3-ITD positive AML. While conditional gene targeting by Cre-recombinase is a powerful technology that has revolutionized biomedical research, consequences of Cre-expression such as lack of fidelity, toxicity or off-target effects need to be taken into consideration. We report on a transgenic murine model of FLT3-ITD induced disease, where Cre-recombinase expression alone, and in the absence of a conditional allele, gives rise to an aggressive leukemia phenotype. Here, expression of various Cre-recombinases leads to polyclonal expansion of FLT3ITD/ITD progenitor cells, induction of a differentiation block and activation of Myc-dependent gene expression programs. Our report is intended to alert the scientific community of potential risks associated with using this specific mouse model and of unexpected effects of Cre-expression when investigating cooperative oncogenic mutations in murine models of cancer.

2023-02-08 | GSE212221 | GEO

Cre-recombinase expression cooperates with FLT3ITD/ITD to induce acute myeloid leukemia [RNASeq_SclCre_JQ1_vs_vehicle]

2023-02-08 | GSE212220 | GEO

Cre-recombinase expression cooperates with FLT3ITD/ITD to induce acute myeloid leukemia [RNASeq_Mx1Cre_BM]

2023-02-08 | GSE212216 | GEO