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Generation of human endometrial knockout cell lines with the CRISPR/Cas9 system confirms the prostaglandin F2alpha synthase activity of aldo-ketoreductase 1B1

Lacroix Pepin, N.; Chapdelaine, P.; Rodriguez, Y.; Tremblay, J.P.; Fortier, M.A.; Mol. Hum. Reprod. 20, 650-663 (2014)

Data extracted from this reference:

Application
EC Number
Application
Commentary
Organism
1.1.1.188
molecular biology
genome editing of human cell lines can be used to complement mouse KO models to validate the function of genes in differentiated tissues and cells
Homo sapiens
Cloned(Commentary)
EC Number
Commentary
Organism
1.1.1.188
gene AKR1B1, cloning from genomic DNA, DNA and amino acid sequence determination and analysis, recombinant expression in immortalized human endometrial stromal cells, HIESC-2 cells, expression analysis in clones 16-1, 16-2, and 16-4. Clones 16-1 and 16-2 present with a complete absence of AKR1B1 protein, while clone 16-4 is able to express the AKR1B1 protein. Following treatment of cells with interleukin-1beta, both wild-type and clone 16-4 cells exhibit a similar response, but clone 16-2 does not respond with increased production of eitherPGF2alpha or PGE2. At the protein level, IL-1beta induces an increase in mPGES-1 and COX-2 protein expression but in clone 16-2 the relative increase of Cox-2 protein is weaker
Homo sapiens
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.188
additional information
knockout of AKR1B1 gene expression in human endometrial cell lines using the CRISPR/Cas9 system. Clone 16-2 exhibits deletion of 68 and 2 nucleotides, respectively, on each of the alleles. Cells from this clone lose their ability to produce PGF2alpha but maintain their original human endometrial stromal cells-2 phenotype including the capacity to decidualize in the presence of progesterone (medroxyprogesterone acetate) and 8-bromo-cAMP. Knockout cells also maintain their ability to increase PGE2 production in response to interleukin-1beta
Homo sapiens
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.188
prostaglandin H2 + NADP+
Homo sapiens
-
prostaglandin F2alpha + NADPH + H+
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.188
Homo sapiens
P15121
-
-
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
1.1.1.188
uterine endometrium
-
Homo sapiens
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.188
prostaglandin H2 + NADP+
-
741012
Homo sapiens
prostaglandin F2alpha + NADPH + H+
-
-
-
?
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.188
NADP+
-
Homo sapiens
Application (protein specific)
EC Number
Application
Commentary
Organism
1.1.1.188
molecular biology
genome editing of human cell lines can be used to complement mouse KO models to validate the function of genes in differentiated tissues and cells
Homo sapiens
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.188
gene AKR1B1, cloning from genomic DNA, DNA and amino acid sequence determination and analysis, recombinant expression in immortalized human endometrial stromal cells, HIESC-2 cells, expression analysis in clones 16-1, 16-2, and 16-4. Clones 16-1 and 16-2 present with a complete absence of AKR1B1 protein, while clone 16-4 is able to express the AKR1B1 protein. Following treatment of cells with interleukin-1beta, both wild-type and clone 16-4 cells exhibit a similar response, but clone 16-2 does not respond with increased production of eitherPGF2alpha or PGE2. At the protein level, IL-1beta induces an increase in mPGES-1 and COX-2 protein expression but in clone 16-2 the relative increase of Cox-2 protein is weaker
Homo sapiens
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.188
NADP+
-
Homo sapiens
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.188
additional information
knockout of AKR1B1 gene expression in human endometrial cell lines using the CRISPR/Cas9 system. Clone 16-2 exhibits deletion of 68 and 2 nucleotides, respectively, on each of the alleles. Cells from this clone lose their ability to produce PGF2alpha but maintain their original human endometrial stromal cells-2 phenotype including the capacity to decidualize in the presence of progesterone (medroxyprogesterone acetate) and 8-bromo-cAMP. Knockout cells also maintain their ability to increase PGE2 production in response to interleukin-1beta
Homo sapiens
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.188
prostaglandin H2 + NADP+
Homo sapiens
-
prostaglandin F2alpha + NADPH + H+
-
-
?
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
1.1.1.188
uterine endometrium
-
Homo sapiens
-
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.188
prostaglandin H2 + NADP+
-
741012
Homo sapiens
prostaglandin F2alpha + NADPH + H+
-
-
-
?
General Information
EC Number
General Information
Commentary
Organism
1.1.1.188
evolution
most prostaglandin F2alpha synthases (PGFS) identified to date are aldo-ketoreductases, AKRs
Homo sapiens
1.1.1.188
metabolism
AKR1B1 is involved in the synthesis of PGF2alpha. Pathways of prostaglandin F2alpha biosynthesis in human cells, overview
Homo sapiens
1.1.1.188
physiological function
prostaglandins (PGs) are important regulators of female reproductive function. The primary PGs produced in the endometrium are PGE2 and PGF2alpha. Role of aldo-ketoreductase (AKR)1B1 in increased PGF2alpha production by human endometrial cells following stimulation with interleukin-1beta (IL-1beta)
Homo sapiens
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.1.1.188
evolution
most prostaglandin F2alpha synthases (PGFS) identified to date are aldo-ketoreductases, AKRs
Homo sapiens
1.1.1.188
metabolism
AKR1B1 is involved in the synthesis of PGF2alpha. Pathways of prostaglandin F2alpha biosynthesis in human cells, overview
Homo sapiens
1.1.1.188
physiological function
prostaglandins (PGs) are important regulators of female reproductive function. The primary PGs produced in the endometrium are PGE2 and PGF2alpha. Role of aldo-ketoreductase (AKR)1B1 in increased PGF2alpha production by human endometrial cells following stimulation with interleukin-1beta (IL-1beta)
Homo sapiens