Dgcr8 knockout approaches to understand microRNA functions in vitro and in vivo

Abstract

Biologic function of the majority of microRNAs (miRNAs) is still unknown. Uncovering the function of miRNAs is hurdled by redundancy among different miRNAs. The deletion of Dgcr8 leads to the deficiency in producing all canonical miRNAs, therefore, overcoming the redundancy issue. Dgcr8 knockout strategy has been instrumental in understanding the function of miRNAs in a variety of cells in vitro and in vivo. In this review, we will first give a brief introduction about miRNAs, miRNA biogenesis pathway and the role of Dgcr8 in miRNA biogenesis. We will then summarize studies performed with Dgcr8 knockout cell models with a focus on embryonic stem cells. After that, we will summarize results from various in vivo Dgcr8 knockout models. Given significant phenotypic differences in various tissues between Dgcr8 and Dicer knockout, we will also briefly review current progresses on understanding miRNA-independent functions of miRNA biogenesis factors. Finally, we will discuss the potential use of a new strategy to stably express miRNAs in Dgcr8 knockout cells. In future, Dgcr8 knockout approaches coupled with innovations in miRNA rescue strategy may provide further insights into miRNA functions in vitro and in vivo.

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Change history

  • 29 April 2019

    The section: “miRNA‑independent functions of DICER” was missed between the section “miRNA‑independent functions of DROSHA and DGCR8” and the section “The Dgcr8 knockout strategy to study miRNA functions” in the original publications.

Abbreviations

APA:

Alternative polyadenylation

AGO:

Argonaute

cKO:

Conditional knockout

cNCCs:

Cardiac neural crest cells

COX-2:

Cyclooxygenase 2

cTECs:

Cortical thymic epithelial cells

DGCR:

DiGeorge syndrome chromosomal (or critical) region

DISME:

DGCR8-independent stable miRNA expression strategy

EMT:

Epithelial–mesenchymal transition

EpiLC:

Epiblast-like cells

EpiSCs:

Epiblast stem cells

ESC:

Embryonic stem cell

ESCC:

ESC-specific cell cycle regulating

GPCs:

Glia progenitor cells

iKO:

Inducible knockout

iPSCs:

Induced pluripotent stem cells

LINE-1:

Long interspersed element 1

lncRNA:

Long noncoding RNA

MC:

Microprocessor complex

miRNA:

microRNA

mTECs:

Medullary thymic epithelial cells

NK:

Natural killer

NSCs:

Neural stem cells

PACT:

Protein activator of PKR

PTA:

Persistent truncus arteriosus

RISC:

RNA-induced silencing complex

RPE:

Retinal pigmented epithelium

rRNA:

Ribosomal RNA

SCs:

Schwann cells

Shh:

Sonic hedgehog

shRNA:

Short hairpin RNA

snoRNA:

Small nucleolar RNA

TDP43:

TAR DNA-binding protein 43

TECs:

Thymic epithelial cells

TRBP:

HIV trans-activating response RNA-binding protein

Th cells:

Helper T cells

tRNA:

Transfer RNA

VSD:

Ventricular septal defect

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Acknowledgements

The research in Wang laboratory is supported by the National Key Research and Development Program of China (2016YFA0100701 and 2018YFA0107601) and the National Natural Science Foundation of China (31471222, 31622033, 31821091 and 91640116). WTG is supported by the Fundamental Research Funds for the Central Universities (3332018008). Due to the breadth of this review, we apologize for the unavoidable exclusion of references to research done by many outstanding investigators working in relevant areas.

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Guo, W., Wang, Y. Dgcr8 knockout approaches to understand microRNA functions in vitro and in vivo. Cell. Mol. Life Sci. 76, 1697–1711 (2019). https://doi.org/10.1007/s00018-019-03020-9

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Keywords

  • Drosha
  • Cell cycle
  • Glycolysis
  • Alternative splicing
  • Glial progenitor cells
  • Reproductive system
  • Neural system
  • Immune system