ArticleBiochemical and biophysical research communications2017
Polycistronic tRNA and CRISPR guide-RNA enables highly efficient multiplexed genome engineering in human cells.
Article in Biochemical and biophysical research communications, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 45 papers.
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Who cites it
45 citing papers in PubMed, 66 citations in OpenAlex.
- Comprehensive genome editing and signaling pathway engineering evolve potent and safe adenoviral producer cell.Communications biology · 2026Article
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- Targeted multiplex gene knockouts in Lemna minor using CRISPR/Cas9.Transgenic research · 2026Article
- Phase-Separation of YAP Mediates AJUBA Super Enhancer Activation to Promote Aberrant Mitosis in Breast Cancer.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- CRISPR-mediated conditional mutagenesis ofbioRxiv : the preprint server for biology · 2026Article
- A next-generation dual guide CRISPR system for genetic interaction library screening.Nature communications · 2025Article
- Anticodon Engineered Transfer RNA (tRNAAdvanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Improving the Efficiency of CRISPR/Cas9-Mediated Non-Homologous End Joining Gene Knockout Using Small Molecules in Porcine Cells.Biomolecules · 2025Article
- Innovations in Cattle Breeding Technology: Prospects in the Era of Gene Editing.Animals : an open access journal from MDPI · 2025Review
- Application of a multiplex CRISPR/Cas9 strategy for elimination of selection markers from transgenic plants.Frontiers in genome editing · 2025Article
- Efficient genome editing in medaka (Journal of Zhejiang University. Science. B · 2024Article
- Review
- How to use CRISPR/Cas9 in plants: from target site selection to DNA repair.Journal of experimental botany · 2024Review
- The application of CRISPR-Cas system inHeliyon · 2024Article
- Article
- Base editing enables duplex point mutagenesis inFrontiers in bioengineering and biotechnology · 2023Article
- Epigenetic modifications as therapeutic targets in atherosclerosis: a focus on DNA methylation and non-coding RNAs.Frontiers in cardiovascular medicine · 2023Review
- Maximizing CRISPRi efficacy and accessibility with dual-sgRNA libraries and optimal effectors.eLife · 2022Article
- Efficient modification and preparation of circular DNA for expression in cell culture.Communications biology · 2022Article
- Multiplex base- and prime-editing with drive-and-process CRISPR arrays.Nature communications · 2022Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 2 countries.
Funding
Abstract
CRISPR/Cas9 has been widely used for genomic editing in many organisms. Many human diseases are caused by multiple mutations. The CRISPR/Cas9 system provides a potential tool to introduce multiple mutations in a genome. To mimic complicated genomic variants in human diseases, such as multiple gene deletions or mutations, two or more small guide RNAs (sgRNAs) need to be introduced all together. This can be achieved by separate Pol III promoters in a construct. However, limited enzyme sites and increased insertion size lower the efficiency to make a construct. Here, we report a strategy to quickly assembly multiple sgRNAs in one construct using a polycistronic-tRNA-gRNA (PTG) strategy. Taking advantage of the endogenous tRNA processing system in mammalian cells, we efficiently express multiple sgRNAs driven using only one Pol III promoter. Using an all-in-one construct carrying PTG, we disrupt the deacetylase domain in multiple histone deacetylases (HDACs) in human cells simultaneously. We demonstrate that multiple HDAC deletions significantly affect the activation of the Wnt-signaling pathway. Thus, this method enables to efficiently target multiple genes and provide a useful tool to establish mutated cells mimicking human diseases.
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Registered trials
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.