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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

In situ hybridization on E18.5 miR-142-/- embryonic lung sections.In situ hybridization for miR-142-3p (A-D) and miR-142-5p (E-H) showing significant reduction of the corresponding expression levels in miR-142-/- compared to WT E18.5 embryonic lung sections. (I-J). Quantification of the in situ hybridization signals indicating a strong reduction of both isoforms. Scale bar: (A,C,E,G): 20 μm and (B,D,F,H): 4 μm.
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pone.0136913.g004: In situ hybridization on E18.5 miR-142-/- embryonic lung sections.In situ hybridization for miR-142-3p (A-D) and miR-142-5p (E-H) showing significant reduction of the corresponding expression levels in miR-142-/- compared to WT E18.5 embryonic lung sections. (I-J). Quantification of the in situ hybridization signals indicating a strong reduction of both isoforms. Scale bar: (A,C,E,G): 20 μm and (B,D,F,H): 4 μm.

Mentions: Using specific digoxygenin-labeled probes for these two microRNAs, in-situ hybridization showed that miR-142-3p and miR-142-5p are expressed in both the mesenchyme and the epithelium of the E18.5 lung (Fig 4A, 4B, 4E and 4F). The level of expression of both microRNAs was significantly decreased in miR142- samples compared to WT samples (Fig 4C, 4D, 4G, 4H vs. 4A, 4B, 4E and 4F). Blinded quantification of the expression of the respective isoforms of miR-142 confirms the very low level of expression of both miR-142 isoforms (Fig 4I and 4J).


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)

In situ hybridization on E18.5 miR-142-/- embryonic lung sections.In situ hybridization for miR-142-3p (A-D) and miR-142-5p (E-H) showing significant reduction of the corresponding expression levels in miR-142-/- compared to WT E18.5 embryonic lung sections. (I-J). Quantification of the in situ hybridization signals indicating a strong reduction of both isoforms. Scale bar: (A,C,E,G): 20 μm and (B,D,F,H): 4 μm.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0136913.g004: In situ hybridization on E18.5 miR-142-/- embryonic lung sections.In situ hybridization for miR-142-3p (A-D) and miR-142-5p (E-H) showing significant reduction of the corresponding expression levels in miR-142-/- compared to WT E18.5 embryonic lung sections. (I-J). Quantification of the in situ hybridization signals indicating a strong reduction of both isoforms. Scale bar: (A,C,E,G): 20 μm and (B,D,F,H): 4 μm.
Mentions: Using specific digoxygenin-labeled probes for these two microRNAs, in-situ hybridization showed that miR-142-3p and miR-142-5p are expressed in both the mesenchyme and the epithelium of the E18.5 lung (Fig 4A, 4B, 4E and 4F). The level of expression of both microRNAs was significantly decreased in miR142- samples compared to WT samples (Fig 4C, 4D, 4G, 4H vs. 4A, 4B, 4E and 4F). Blinded quantification of the expression of the respective isoforms of miR-142 confirms the very low level of expression of both miR-142 isoforms (Fig 4I and 4J).

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