ArticlePloS one2018
One step generation of customizable gRNA vectors for multiplex CRISPR approaches through string assembly gRNA cloning (STAgR).
Article in PloS one, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed.
- RAPID-DASH: fast and efficient assembly of guide RNA arrays for multiplexed CRISPR-Cas9 applications.Synthetic biology (Oxford, England) · 2026Article
- RAPID-DASH: Fast and Efficient Assembly of Guide RNA Arrays for Multiplexed CRISPR-Cas9 Applications.bioRxiv : the preprint server for biology · 2025Article
- Efficient DNA- and virus-free engineering of cellular transcriptomic states using dCas9 ribonucleoprotein (dRNP) complexes.Nucleic acids research · 2025Article
- High-accuracy crRNA array assembly strategy for multiplex CRISPR.Molecular therapy. Nucleic acids · 2025Article
- Genetic manipulations of nonmodel gut microbes.iMeta · 2024Article
- Therapeutic targeting Tudor domains in leukemia via CRISPR-Scan Assisted Drug Discovery.Science advances · 2024Article
- Parkinson's disease motor symptoms rescue by CRISPRa-reprogramming astrocytes into GABAergic neurons.EMBO molecular medicine · 2022Article
- Multiplexed Genome Editing for Efficient Phenotypic Screening in Zebrafish.Veterinary sciences · 2022Article
- Article
- New Therapies to Correct the Cystic Fibrosis Basic Defect.International journal of molecular sciences · 2021Review
- CRISPR-Mediated Induction of Neuron-Enriched Mitochondrial Proteins Boosts Direct Glia-to-Neuron Conversion.Cell stem cell · 2021Article
- MARK2 phosphorylates eIF2α in response to proteotoxic stress.PLoS biology · 2021Article
- Non-codon Optimized PiggyBac Transposase Induces Developmental Brain Aberrations: A Call forFrontiers in cell and developmental biology · 2021Article
- FAMSi: A Synthetic Biology Approach to the Fast Assembly of Multiplex siRNAs for Silencing Gene Expression in Mammalian Cells.Molecular therapy. Nucleic acids · 2020Article
- Multiplexed CRISPR technologies for gene editing and transcriptional regulation.Nature communications · 2020Review
- Developing a Versatile Shotgun Cloning Strategy for Single-Vector-Based Multiplex Expression of Short Interfering RNAs (siRNAs) in Mammalian Cells.ACS synthetic biology · 2019Article
- Targeted removal of epigenetic barriers during transcriptional reprogramming.Nature communications · 2019Article
- A novel cloning strategy for one-step assembly of multiplex CRISPR vectors.Scientific reports · 2018Article
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9 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Novel applications based on the bacterial CRISPR system make genetic, genomic, transcriptional and epigenomic engineering widely accessible for the first time. A significant advantage of CRISPR over previous methods is its tremendous adaptability due to its bipartite nature. Cas9 or its engineered variants define the molecular effect, while short gRNAs determine the targeting sites. A majority of CRISPR approaches depend on the simultaneous delivery of multiple gRNAs into single cells, either as an essential precondition, to increase responsive cell populations or to enhance phenotypic outcomes. Despite these requirements, methods allowing the efficient generation and delivery of multiple gRNA expression units into single cells are still sparse. Here we present STAgR (String assembly gRNA cloning), a single step gRNA multiplexing system, that obtains its advantages by employing the N20 targeting sequences as necessary homologies for Gibson assembly. We show that STAgR allows reliable and cost-effective generation of vectors with high numbers of gRNAs enabling multiplexed CRISPR approaches. Moreover, STAgR is easily customizable, as vector backbones as well as gRNA structures, numbers and promoters can be freely chosen and combined. Finally, we demonstrate STAgR's widespread functionality, its efficiency in multi-targeting approaches, using it for both, genome and transcriptome editing, as well as applying it in vitro and in vivo.
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