Related Experiment Video
Updated: Jan 26, 2026
Mouse Fetal Liver Culture System to Dissect Target Gene Functions at the Early and Late Stages of Terminal Erythropoiesis
Published on: September 9, 2014
Regulation of gene expression by miR-144/451 during mouse erythropoiesis
Peng Xu1, Lance E Palmer2, Christophe Lechauve1
1Department of Hematology and.
Abstract:
The microRNA (miRNA) locus miR-144/451 is abundantly expressed in erythrocyte precursors, facilitating their terminal maturation and protecting against oxidant stress. However, the full repertoire of erythroid miR-144/451 target messenger RNAs (mRNAs) and associated cellular pathways is unknown. In general, the numbers of mRNAs predicted to be targeted by an miRNA vary greatly from hundreds to thousands, and are dependent on experimental approaches. To comprehensively and accurately identify erythroid miR-144/451 target mRNAs, we compared gene knockout and wild-type fetal liver erythroblasts by RNA sequencing, quantitative proteomics, and RNA immunoprecipitation of Argonaute (Ago), a component of the RNA-induced silencing complex that binds miRNAs complexed to their target mRNAs. Argonaute bound ∼1400 erythroblast mRNAs in a miR-144/451-dependent manner, accounting for one-third of all Ago-bound mRNAs. However, only ∼100 mRNAs were stabilized after miR-144/451 loss. Thus, miR-144 and miR-451 deregulate <10% of mRNAs that they bind, a characteristic that likely applies generally to other miRNAs. Using stringent selection criteria, we identified 53 novel miR-144/451 target mRNAs. One of these, Cox10, facilitates the assembly of mitochondrial electron transport complex IV. Loss of miR-144/451 caused increased Cox10 mRNA and protein, accumulation of complex IV, and increased mitochondrial membrane potential with no change in mitochondrial mass. Thus, miR-144/451 represses mitochondrial respiration during erythropoiesis by inhibiting the production of Cox10.
Insights
MicroRNA miR-144/451 is crucial for red blood cell development. This study identifies novel targets, revealing its role in repressing mitochondrial respiration by inhibiting Cox10 production during erythropoiesis.
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- The microRNA (miRNA) locus miR-144/451 plays a key role in erythrocyte maturation and protection against oxidative stress.
- The complete set of target messenger RNAs (mRNAs) and cellular pathways regulated by erythroid miR-144/451 remains largely unknown.
Purpose of the Study:
- To comprehensively identify erythroid miR-144/451 target mRNAs and understand their associated cellular pathways.
- To elucidate the specific role of miR-144/451 in regulating mitochondrial function during erythropoiesis.
Main Methods:
- Comparative RNA sequencing and quantitative proteomics of gene knockout and wild-type fetal liver erythroblasts.
- RNA immunoprecipitation of Argonaute (Ago) to identify miRNA-bound mRNAs.
- Stringent selection criteria to validate novel miRNA targets.
Main Results:
- Argonaute bound approximately 1400 erythroblast mRNAs in a miR-144/451-dependent manner.
- Identified 53 novel miR-144/451 target mRNAs, including Cox10, which is involved in mitochondrial electron transport complex IV assembly.
- Loss of miR-144/451 led to increased Cox10 expression, complex IV accumulation, and elevated mitochondrial membrane potential, indicating repressed mitochondrial respiration.
Conclusions:
- miR-144/451 deregulates a small fraction (<10%) of the mRNAs it binds, a characteristic potentially common to other miRNAs.
- miR-144/451 represses mitochondrial respiration during erythropoiesis by inhibiting Cox10 production.
- This study provides novel insights into miRNA-mediated regulation of mitochondrial function in red blood cell development.
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