ArticleMolecular cell2018
Rapid and Scalable Characterization of CRISPR Technologies Using an E. coli Cell-Free Transcription-Translation System.
Article in Molecular cell, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 86 papers.
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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
86 citing papers in PubMed, 210 citations in OpenAlex.
- Expanding the Microbial Genomic Landscape and Biotechnological Applications of CRISPR-Cas Systems.Biology · 2026Review
- GenomePAM directs PAM characterization and engineering of CRISPR-Cas nucleases using mammalian genome repeats.Nature biomedical engineering · 2026Article
- Article
- PAM-interacting domain turn-helix 51 motifs can improve Cas9-SpRY activity.Nucleic acids research · 2025Article
- Efficient encapsulation of CRISPR-Cas9 RNP in bioreducible nanogels and release in a cytosol-mimicking environment.Discover nano · 2025Article
- GenomePAM directs PAM characterization and engineering of CRISPR-Cas nucleases using mammalian genome repeats.Research square · 2025Article
- Responses of natural plastisphere community and zooplankton to microplastic pollution: a review on novel remediation strategies.Archives of microbiology · 2025Review
- Article
- Self-growing protocell models in aqueous two-phase system induced by internal DNA replication reaction.Nature communications · 2025Article
- Cell-free expression system: a promising platform for bacteriophage production and engineering.Microbial cell factories · 2025Review
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- A catalog of gene editing sites and genetic variations in editing sites in model organisms.BMC genomics · 2024Article
- Review
- Ten Years of the Synthetic Biology Summer Course at Cold Spring Harbor Laboratory.ACS synthetic biology · 2024Review
- Characterization ofmSphere · 2024Article
- TracrRNA reprogramming enables direct PAM-independent detection of RNA with diverse DNA-targeting Cas12 nucleases.Nature communications · 2024Article
- Reversible Control of Gene Expression by Guest-Modified Adenosines in a Cell-Free System via Host-Guest Interaction.Journal of the American Chemical Society · 2024Article
- Phylogenomics and genetic analysis of solvent-producing Clostridium species.Scientific data · 2024Article
- CRISPR-based screening of small RNA modulators of bile susceptibility inProceedings of the National Academy of Sciences of the United States of America · 2024Article
- Interrogating two extensively self-targeting Type I CRISPR-Cas systems in Xanthomonas albilineans reveals distinct anti-CRISPR proteins that block DNA degradation.Nucleic acids research · 2024Article
26 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
12 authors at 3 institutions in 1 country.
Funding
Abstract
CRISPR-Cas systems offer versatile technologies for genome engineering, yet their implementation has been outpaced by ongoing discoveries of new Cas nucleases and anti-CRISPR proteins. Here, we present the use of E. coli cell-free transcription-translation (TXTL) systems to vastly improve the speed and scalability of CRISPR characterization and validation. TXTL can express active CRISPR machinery from added plasmids and linear DNA, and TXTL can output quantitative dynamics of DNA cleavage and gene repression-all without protein purification or live cells. We used TXTL to measure the dynamics of DNA cleavage and gene repression for single- and multi-effector CRISPR nucleases, predict gene repression strength in E. coli, determine the specificities of 24 diverse anti-CRISPR proteins, and develop a fast and scalable screen for protospacer-adjacent motifs that was successfully applied to five uncharacterized Cpf1 nucleases. These examples underscore how TXTL can facilitate the characterization and application of CRISPR technologies across their many uses.
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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.