ArticleRNA (New York, N.Y.)2014
A TALEN-based strategy for efficient bi-allelic miRNA ablation in human cells.
Article in RNA (New York, N.Y.), 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 23 citations in OpenAlex.
- Hijacking exosome biogenesis: viral glycoproteins as modular scaffolds for engineering functionalized extracellular vesicles.Nanoscale · 2026Article
- MiRNA fine tuning for crop improvement: using advance computational models and biotechnological tools.Molecular biology reports · 2022Review
- Recapture Lysosomal Enzyme Deficiency via Targeted Gene Disruption in the Human Near-Haploid Cell Line HAP1.Genes · 2021Article
- Disruption of miRNA sequences by TALENs and CRISPR/Cas9 induces varied lengths of miRNA production.Plant biotechnology journal · 2020Article
- TargetLink, a new method for identifying the endogenous target set of a specific microRNA in intact living cells.RNA biology · 2017Article
- Knockout of miR-221 and miR-222 reveals common and specific targets for paralogous miRNAs.RNA biology · 2017Article
- Pseudotyping exosomes for enhanced protein delivery in mammalian cells.International journal of nanomedicine · 2017Article
- AAV-based dual-reporter circuit for monitoring cell signaling in living human cells.Journal of biological engineering · 2017Article
- Viral Cre-LoxP tools aid genome engineering in mammalian cells.Journal of biological engineering · 2017Article
- MicroRNAs as regulatory elements in immune system logic.Nature reviews. Immunology · 2016Review
- An Optimized Protocol for Packaging Pseudotyped Integrase Defective Lentivirus.Biological procedures online · 2016Article
- Functional screen reveals essential roles of miR-27a/24 in differentiation of embryonic stem cells.The EMBO journal · 2015Article
- Disruption of microRNA-21 by TALEN leads to diminished cell transformation and increased expression of cell-environment interaction genes.Cancer letters · 2015Article
- Gene therapy for cancer: present status and future perspective.Molecular and cellular therapies · 2014Review
- Tiny giants of gene regulation: experimental strategies for microRNA functional studies.Wiley interdisciplinary reviews. Developmental biologyReview
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Significant progress in the functional understanding of microRNAs (miRNAs) has been made in mice, but a need remains to develop efficient tools for bi-allelic knockouts of miRNA in the human genome. Transcription activator-like effector nucleases (TALENs) provide an exciting platform for targeted gene ablation in cultured human cells, but bi-allelic modifications induced by TALENs alone occur at low frequency, making screening for double knockouts a tedious task. Here, we present an approach that is highly efficient in bi-allelic miRNA ablation in the human genome by combining TALENs targeting to the miRNA seed region with a homologous recombination donor vector and a positive selection strategy. A pilot test of this approach demonstrates bi-allelic miR-21 gene disruption at high frequency (∼87%) in cultured HEK293 cells. Analysis of three independent clones showed a total loss of miR-21 expression. Phenotypical analysis revealed increased miR-21 target gene expression, reduced cell proliferation, and alterations of global miRNA expression profiles. Taken together, our study reveals a feasible and efficient approach for bi-allelic miRNA ablation in cultured human cells and demonstrates its usefulness in elucidating miRNA function in human cells.
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