ArticlePlants (Basel, Switzerland)2021
Construction of Multiple Guide RNAs in CRISPR/Cas9 Vector Using Stepwise or Simultaneous Golden Gate Cloning: Case Study for Targeting the
Article in Plants (Basel, Switzerland), 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
13 citing papers in PubMed, 15 citations in OpenAlex.
- Multiplex FAD2/FATB Editing Generates Ultra-High-Oleic, Low-Saturate Soybean With Increased Seed Fatty Acid Content.Plant biotechnology journal · 2026Article
- Cell wall invertase improves grain nutrition via regulating sugar and hormone metabolism gene expression in transgenic soybean.Annals of botany · 2026Article
- CRISPR/Cas9 in soybean research: a measured leap toward genetic refinement.Frontiers in genome editing · 2026Review
- BvHP4b gene in red beet promotes tuber enlargement and enhances resistance to Pst DC3000.BMC genomics · 2025Article
- CRISPR/Cas genome editing in soybean: challenges and new insights to overcome existing bottlenecks.Journal of advanced research · 2025Review
- Applying conventional and cell-type-specific CRISPR/Cas9 genome editing in legume plants.aBIOTECH · 2025Review
- How to use CRISPR/Cas9 in plants: from target site selection to DNA repair.Journal of experimental botany · 2024Review
- SlTrxh functions downstream of SlMYB86 and positively regulates nitrate stress tolerance via S-nitrosation in tomato seedling.Horticulture research · 2024Article
- Genetic manipulations of nonmodel gut microbes.iMeta · 2024Article
- Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding.Plants (Basel, Switzerland) · 2023Review
- Development of Ethyl Methanesulfonate Mutant Edamame Soybean (Plants (Basel, Switzerland) · 2022Article
- CRISPR/Cas9 in Planta Hairy Root Transformation: A Powerful Platform for Functional Analysis of Root Traits in Soybean.Plants (Basel, Switzerland) · 2022Review
- Applications and prospects of genome editing in plant fatty acid and triacylglycerol biosynthesis.Frontiers in plant science · 2022Review
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
CRISPR/Cas9 is a commonly used technique in reverse-genetics research to knock out a gene of interest. However, when targeting a multigene family or multiple genes, it is necessary to construct a vector with multiple single guide RNAs (sgRNAs) that can navigate the Cas9 protein to the target site. In this protocol, the Golden Gate cloning method was used to generate multiple sgRNAs in the Cas9 vector. The vectors used were pHEE401E_UBQ_Bar and pBAtC_tRNA, which employ a one-promoter/one-sgRNA and a polycistronic-tRNA-gRNA strategy, respectively. Golden Gate cloning was performed with type IIS restriction enzymes to generate gRNA polymers for vector inserts. Four sgRNAs containing the pHEE401E_UBQ_Bar vector and four to six sgRNAs containing the pBAtC_tRNA vector were constructed. In practice, we constructed multiple sgRNAs targeting multiple genes of
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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.