ArticleStem cell reports2016
Efficient CRISPR/Cas9-Mediated Versatile, Predictable, and Donor-Free Gene Knockout in Human Pluripotent Stem Cells.
Article in Stem cell reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed, 46 citations in OpenAlex.
- Optimization of gene knockout approaches and sgRNA selection in hPSCs with inducible Cas9 expression.Scientific reports · 2025Article
- Early blood immune molecular alterations in cynomolgus monkeys with a PSEN1 mutation causing familial Alzheimer's disease.Alzheimer's & dementia : the journal of the Alzheimer's Association · 2024Article
- Initial Characterization of WDR5B Reveals a Role in the Proliferation of Retinal Pigment Epithelial Cells.Cells · 2024Article
- Functional annotation of variants of the BRCA2 gene via locally haploid human pluripotent stem cells.Nature biomedical engineering · 2024Article
- Pioneer factor ASCL1 cooperates with the mSWI/SNF complex at distal regulatory elements to regulate human neural differentiation.Genes & development · 2023Article
- Efficient deletion of microRNAs using CRISPR/Cas9 with dual guide RNAs.Frontiers in molecular biosciences · 2023Article
- Robust genome editing via modRNA-based Cas9 or base editor in human pluripotent stem cells.Cell reports methods · 2022Article
- Functional annotation of lncRNA in high-throughput screening.Essays in biochemistry · 2021Review
- Toward precise CRISPR DNA fragment editing and predictable 3D genome engineering.Journal of molecular cell biology · 2021Review
- Mapping germ-layer specification preventing genes in hPSCs via genome-scale CRISPR screening.iScience · 2021Article
- Avian Toll-like receptor 3 isoforms and evaluation of Toll-like receptor 3-mediated immune responses using knockout quail fibroblast cells.Poultry science · 2020Article
- Novel Approaches to Profile Functional Long Noncoding RNAs Associated with Stem Cell Pluripotency.Current genomics · 2020Review
- SOX21 Ensures Rostral Forebrain Identity by Suppression of WNT8B during Neural Regionalization of Human Embryonic Stem Cells.Stem cell reports · 2019Article
- Methods and applications of CRISPR/Cas system for genome editing in stem cells.Cell regeneration (London, England) · 2019Review
- High-throughput micropatterning platform reveals Nodal-dependent bisection of peri-gastrulation-associated versus preneurulation-associated fate patterning.PLoS biology · 2019Article
- A Brief Overview of lncRNAs in Endothelial Dysfunction-Associated Diseases: From Discovery to Characterization.Epigenomes · 2019Review
- Transcriptionally active HERV-H retrotransposons demarcate topologically associating domains in human pluripotent stem cells.Nature genetics · 2019Article
- WNT/NOTCH Pathway Is Essential for the Maintenance and Expansion of Human MGE Progenitors.Stem cell reports · 2019Article
- Genetic Engineering of Human Embryonic Stem Cells for Precise Cell Fate Tracing during Human Lineage Development.Stem cell reports · 2018Article
- Sin3a-Tet1 interaction activates gene transcription and is required for embryonic stem cell pluripotency.Nucleic acids research · 2018Article
Corrections and comments
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Authors and funding
16 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Loss-of-function studies in human pluripotent stem cells (hPSCs) require efficient methodologies for lesion of genes of interest. Here, we introduce a donor-free paired gRNA-guided CRISPR/Cas9 knockout strategy (paired-KO) for efficient and rapid gene ablation in hPSCs. Through paired-KO, we succeeded in targeting all genes of interest with high biallelic targeting efficiencies. More importantly, during paired-KO, the cleaved DNA was repaired mostly through direct end joining without insertions/deletions (precise ligation), and thus makes the lesion product predictable. The paired-KO remained highly efficient for one-step targeting of multiple genes and was also efficient for targeting of microRNA, while for long non-coding RNA over 8 kb, cleavage of a short fragment of the core promoter region was sufficient to eradicate downstream gene transcription. This work suggests that the paired-KO strategy is a simple and robust system for loss-of-function studies for both coding and non-coding genes in hPSCs.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.