ArticleNature communications2021
Engineering a PAM-flexible SpdCas9 variant as a universal gene repressor.
Article in Nature communications, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
What it found
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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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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
21 citing papers in PubMed, 33 citations in OpenAlex.
- Engineering genetic circuits for dynamic control of central metabolism.Metabolic engineering · 2025Article
- Structural basis of a dual-function type II-B CRISPR-Cas9.Nucleic acids research · 2025Article
- Antibiotic-ResistantMicroorganisms · 2025Review
- CRISPR/Cas9: a sustainable technology to enhance climate resilience in major Staple Crops.Frontiers in genome editing · 2025Review
- Next-generation CRISPR technology for genome, epigenome and mitochondrial editing.Transgenic research · 2024Review
- CRISPR-Cas9-based genome-editing technologies in engineering bacteria for the production of plant-derived terpenoids.Engineering microbiology · 2024Review
- Investigating and Engineering an 1,2-Propanediol-Responsive Transcription Factor-Based Biosensor.ACS synthetic biology · 2024Article
- High-Throughput Screening of PAM-Flexible Cas9 Variants for Expanded Genome Editing in the Silkworm (Insects · 2024Article
- A self-regulated network for dynamically balancing multiple precursors in complex biosynthetic pathways.Metabolic engineering · 2024Article
- Engineering Cas9: next generation of genomic editors.Applied microbiology and biotechnology · 2024Review
- Harnessing non-Watson-Crick's base pairing to enhance CRISPR effectors cleavage activities and enable gene editing in mammalian cells.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- Engineering aACS synthetic biology · 2023Article
- Dynamic Metabolic Control: From the Perspective of Regulation Logic.Synthetic biology and engineering · 2023Article
- Genome editing and cancer therapy: handling the hypoxia-responsive pathway as a promising strategy.Cellular and molecular life sciences : CMLS · 2023Review
- Exploring and engineering PAM-diverse Streptococci Cas9 for PAM-directed bifunctional and titratable gene control in bacteria.Metabolic engineering · 2023Article
- Type I CRISPR-Cas-mediated microbial gene editing and regulation.AIMS microbiology · 2023Review
- Biosensor-assisted titratable CRISPRi high-throughput (BATCH) screening for over-production phenotypes.Metabolic engineering · 2023Article
- Expanding the Scope of Bacterial CRISPR Activation with PAM-Flexible dCas9 Variants.ACS synthetic biology · 2022Article
- Establishing an Autonomous Cascaded Artificial Dynamic (AutoCAD) regulation system for improved pathway performance.Metabolic engineering · 2022Article
- CRISPRactivation-SMS, a message for PAM sequence independent gene up-regulation in Escherichia coli.Nucleic acids research · 2022Article
Corrections and comments
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Authors and funding
9 authors at 1 institution in 1 country.
Funding
Abstract
The RNA-guided CRISPR-associated Cas9 proteins have been widely applied in programmable genome recombination, base editing or gene regulation in both prokaryotes and eukaryotes. SpCas9 from Streptococcus pyogenes is the most extensively engineered Cas9 with robust and manifold functionalities. However, one inherent limitation of SpCas9 is its stringent 5'-NGG-3' PAM requirement that significantly restricts its DNA target range. Here, to repurpose SpCas9 as a universal gene repressor, we generate and screen variants of the deactivated SpCas9 (SpdCas9) with relaxed 5'-CAT-3' PAM compatibility that can bind to the start codon ATG of almost any gene. Stepwise structure-guided mutations of the PAM-interacting residues and auxiliary PAM-proximal residues of the SpdNG (5'-NG-3' PAM) create a PAM-flexible variant SpdNG-LWQT that preferentially accommodates 5'-NRN-3' PAMs. SpdNG-LWQT is demonstrated to be effective in gene repression with the advantage of customizable sgRNA design in both Escherichia coli and Saccharomyces cerevisiae. This work validates the feasibility of purposeful PAM expansion of Cas9 towards signature PAMs and establishes a universal SpdCas9-based gene repressor.
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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.