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CircRNA knockdown based on antisense strategies.

Guillermo Aquino-Jarquin1

  • 1RNA Biology and Genome Editing Section. Genomics, Genetics, and Bioinformatics Research Laboratory. 'Federico Gómez' Children's Hospital of Mexico. Dr. Márquez 162, Doctores, Cuauhtémoc, CP 06720, CDMX, Mexico.

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Summary

Investigating circular RNA (circRNA) functions requires effective knockdown. This review details antisense strategies, including CRISPR-Cas13, for targeting circRNAs to understand their biological roles in cells and animal models.

Keywords:
ASOsAntisense therapyBack-splicingCRISPR-Cas13CRISPR-Cas9CircRNAssiRNA/shRNA

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Circular RNAs (circRNAs) are noncoding RNAs derived from eukaryotic protein-coding genes via back-splicing.
  • Their primary known function is acting as miRNA and protein sponges, but many circRNA functions remain unelucidated.
  • Understanding circRNA biological relevance is crucial for advancing molecular biology.

Purpose of the Study:

  • To review and summarize the feasibility and effectiveness of various antisense strategies for circRNA knockdown.
  • To highlight methods for investigating the biological roles of circRNAs in cellular and animal models.
  • To provide insights into targeting circRNAs for functional genomic studies.

Main Methods:

  • Antisense oligonucleotides (ASOs) for targeted circRNA binding.
  • RNA interference (RNAi) targeting back-splicing junctions.
  • CRISPR-Cas9 for generating circRNA-specific knockouts.
  • CRISPR-Cas13 technology for precise circRNA targeting without affecting host genes.

Main Results:

  • Antisense strategies offer effective means to reduce circRNA levels.
  • CRISPR-Cas13 demonstrates high specificity in targeting circRNAs.
  • These methods enable functional analyses of circRNAs in diverse biological contexts.
  • The review consolidates current knowledge on circRNA knockdown techniques.

Conclusions:

  • CircRNA knockdown via antisense strategies is essential for functional characterization.
  • CRISPR-Cas13 and other antisense approaches provide powerful tools for circRNA research.
  • Further investigation into circRNA functions will advance our understanding of gene regulation and disease.