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Generation and Validation of miR-142 Knock Out Mice.

Shrestha A, Carraro G, El Agha E, Mukhametshina R, Chao CM, Rizvanov A, Barreto G, Bellusci S - PLoS ONE (2015)

Bottom Line: microRNA-142 (miR-142) is an important regulator of many biological processes and associated signaling pathways during embryonic development, homeostasis and disease.The expression of Bzrap1, a gene immediately flanking miR-142 is not altered while the expression of a long non-coding RNA embedded within the miR-142 gene is decreased. miR-142- newborn pups appear normal and are normally represented indicating absence of embryonic lethality.At embryonic day 18.5, miR-142- lungs display increased Wnt signaling associated with the up-regulation of Apc and p300, two previously reported targets of miR-142-3p and -5p, respectively.

View Article: PubMed Central - PubMed

Affiliation: German Center for Lung Research, Excellence Cluster Cardio-Pulmonary System, Universities of Giessen and Marburg Lung Center, Giessen, Hessen, Germany.

ABSTRACT
microRNA-142 (miR-142) is an important regulator of many biological processes and associated signaling pathways during embryonic development, homeostasis and disease. The miR-142 hairpin gives rise to the "guide strand" miR-142-3p and the sister "passenger" strand miR-142-5p. miR-142-3p has been shown to play critical, non-redundant functions in the development of the hematopoietic lineage. We have recently reported that miR-142-3p is critical for the control of Wnt signaling in the mesenchyme of the developing lung. miR-142-5p has been proposed to control adaptive growth in cardiomyocytes postnatally and its increase is associated with extensive apoptosis and cardiac dysfunction in a murine heart failure model. Using homologous recombination, we now report the generation and validation of miR-142- mice. miR-142- mice show a significant decrease in th expression levels of both the 3p and 5p isoforms. The expression of Bzrap1, a gene immediately flanking miR-142 is not altered while the expression of a long non-coding RNA embedded within the miR-142 gene is decreased. miR-142- newborn pups appear normal and are normally represented indicating absence of embryonic lethality. At embryonic day 18.5, miR-142- lungs display increased Wnt signaling associated with the up-regulation of Apc and p300, two previously reported targets of miR-142-3p and -5p, respectively. Adult miR-142- animals display impaired hematopoietic lineage formation identical to previously reported miR-142 gene trap knockdown mice. We report, for the first time, the homologous recombination-based miR-142- mice that will be useful for the scientific community working on the diverse biological functions of miR-142.

No MeSH data available.


Related in: MedlinePlus

Schematic Representation of the miR-142 locus in the mouse genome.miR-142 gene is located in the vicinity of an exon belonging to a LncRNA (A430104N18Rik ENSMUSG00000084796) on chromosome 11. Deletion of miR-142 led to the deletion of the LncRNA exon as well as part of the 5’ UTR of Bzrap1. LncRNA: Long non-coding RNA. Bzrap1: Benzodiazepine receptor (peripheral) associated protein 1.
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pone.0136913.g001: Schematic Representation of the miR-142 locus in the mouse genome.miR-142 gene is located in the vicinity of an exon belonging to a LncRNA (A430104N18Rik ENSMUSG00000084796) on chromosome 11. Deletion of miR-142 led to the deletion of the LncRNA exon as well as part of the 5’ UTR of Bzrap1. LncRNA: Long non-coding RNA. Bzrap1: Benzodiazepine receptor (peripheral) associated protein 1.

Mentions: In mice, the miR-142 gene (ENSMUSG00000065420) is located in chromosome 11 adjacent to the second exon of a long non-coding RNA (A430104N18Rik ENSMUSG00000084796) (Fig 1). The function is still LncRNA is still unknown. Benzodiazepine receptor (peripheral) associated protein 1 (Bzrap1, ENSMUSG00000034156) is also another gene found downstream of miR-142. The miR-142 locus also contains many GC-rich repeat regions, which render PCR amplification and screening difficult. The constitutive and complete deletion of the miR-142 gene was therefore carried out and miR-142 heterozygous mice were generated. Fig 2B shows the genotyping results of E18.5 embryos arising from crossing miR-142 heterozygous mice. Fig 2C shows the validation of our primer sets to amplify either the wild type miR-142 allele (P2/P3), the presence of the LoxP site (P1/P4) or the miR-142- allele (P1/P3).


Generation and Validation of miR-142 Knock Out Mice.

Shrestha A, Carraro G, El Agha E, Mukhametshina R, Chao CM, Rizvanov A, Barreto G, Bellusci S - PLoS ONE (2015)

Schematic Representation of the miR-142 locus in the mouse genome.miR-142 gene is located in the vicinity of an exon belonging to a LncRNA (A430104N18Rik ENSMUSG00000084796) on chromosome 11. Deletion of miR-142 led to the deletion of the LncRNA exon as well as part of the 5’ UTR of Bzrap1. LncRNA: Long non-coding RNA. Bzrap1: Benzodiazepine receptor (peripheral) associated protein 1.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556616&req=5

pone.0136913.g001: Schematic Representation of the miR-142 locus in the mouse genome.miR-142 gene is located in the vicinity of an exon belonging to a LncRNA (A430104N18Rik ENSMUSG00000084796) on chromosome 11. Deletion of miR-142 led to the deletion of the LncRNA exon as well as part of the 5’ UTR of Bzrap1. LncRNA: Long non-coding RNA. Bzrap1: Benzodiazepine receptor (peripheral) associated protein 1.
Mentions: In mice, the miR-142 gene (ENSMUSG00000065420) is located in chromosome 11 adjacent to the second exon of a long non-coding RNA (A430104N18Rik ENSMUSG00000084796) (Fig 1). The function is still LncRNA is still unknown. Benzodiazepine receptor (peripheral) associated protein 1 (Bzrap1, ENSMUSG00000034156) is also another gene found downstream of miR-142. The miR-142 locus also contains many GC-rich repeat regions, which render PCR amplification and screening difficult. The constitutive and complete deletion of the miR-142 gene was therefore carried out and miR-142 heterozygous mice were generated. Fig 2B shows the genotyping results of E18.5 embryos arising from crossing miR-142 heterozygous mice. Fig 2C shows the validation of our primer sets to amplify either the wild type miR-142 allele (P2/P3), the presence of the LoxP site (P1/P4) or the miR-142- allele (P1/P3).

Bottom Line: microRNA-142 (miR-142) is an important regulator of many biological processes and associated signaling pathways during embryonic development, homeostasis and disease.The expression of Bzrap1, a gene immediately flanking miR-142 is not altered while the expression of a long non-coding RNA embedded within the miR-142 gene is decreased. miR-142- newborn pups appear normal and are normally represented indicating absence of embryonic lethality.At embryonic day 18.5, miR-142- lungs display increased Wnt signaling associated with the up-regulation of Apc and p300, two previously reported targets of miR-142-3p and -5p, respectively.

View Article: PubMed Central - PubMed

Affiliation: German Center for Lung Research, Excellence Cluster Cardio-Pulmonary System, Universities of Giessen and Marburg Lung Center, Giessen, Hessen, Germany.

ABSTRACT
microRNA-142 (miR-142) is an important regulator of many biological processes and associated signaling pathways during embryonic development, homeostasis and disease. The miR-142 hairpin gives rise to the "guide strand" miR-142-3p and the sister "passenger" strand miR-142-5p. miR-142-3p has been shown to play critical, non-redundant functions in the development of the hematopoietic lineage. We have recently reported that miR-142-3p is critical for the control of Wnt signaling in the mesenchyme of the developing lung. miR-142-5p has been proposed to control adaptive growth in cardiomyocytes postnatally and its increase is associated with extensive apoptosis and cardiac dysfunction in a murine heart failure model. Using homologous recombination, we now report the generation and validation of miR-142- mice. miR-142- mice show a significant decrease in th expression levels of both the 3p and 5p isoforms. The expression of Bzrap1, a gene immediately flanking miR-142 is not altered while the expression of a long non-coding RNA embedded within the miR-142 gene is decreased. miR-142- newborn pups appear normal and are normally represented indicating absence of embryonic lethality. At embryonic day 18.5, miR-142- lungs display increased Wnt signaling associated with the up-regulation of Apc and p300, two previously reported targets of miR-142-3p and -5p, respectively. Adult miR-142- animals display impaired hematopoietic lineage formation identical to previously reported miR-142 gene trap knockdown mice. We report, for the first time, the homologous recombination-based miR-142- mice that will be useful for the scientific community working on the diverse biological functions of miR-142.

No MeSH data available.


Related in: MedlinePlus