Evidence map›Paper›PMID 32833533›Full record

ArticleThe CRISPR journal2020

Promoter Orientation within an AAV-CRISPR Vector Affects Cas9 Expression and Gene Editing Efficiency.

Lewis E Fry, Caroline F Peddle, Marta Stevanovic, Alun R Barnard, Michelle E McClements, Robert E MacLaren

Open access · greenAbstract read
In one paragraph

Article in The CRISPR journal, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.

0numbers the graph read from it
0cells of the map it votes in
6citing papers in PubMed
0.6field-weighted citation impact, top 35% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

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.

2 · The registry

The trial behind it

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.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

6 citing papers in PubMed, 11 citations in OpenAlex.

  1. Article
  2. Article
  3. Review
  4. CRISPR/Cas9 for hepatitis B virus infection treatment.Immunity, inflammation and disease · 2023
    Review
  5. Building Blocks of Artificial CRISPR-Based Systems beyond Nucleases.International journal of molecular sciences · 2022
    Review
  6. Adenine Base EditingGEN biotechnology · 2022
    Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors at 1 institution in 1 country.

Lewis E FryNuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neurosciences and NIHR Oxford Biomedical Research Centre, University of Oxford, Oxford, United Kingdom; and Oxford University Hospitals NHS Foundation Trust, Oxford, United Kingdom.
Caroline F PeddleNuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neurosciences and NIHR Oxford Biomedical Research Centre, University of Oxford, Oxford, United Kingdom; and Oxford University Hospitals NHS Foundation Trust, Oxford, United Kingdom.
Marta StevanovicNuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neurosciences and NIHR Oxford Biomedical Research Centre, University of Oxford, Oxford, United Kingdom; and Oxford University Hospitals NHS Foundation Trust, Oxford, United Kingdom.
Alun R BarnardNuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neurosciences and NIHR Oxford Biomedical Research Centre, University of Oxford, Oxford, United Kingdom; and Oxford University Hospitals NHS Foundation Trust, Oxford, United Kingdom.
Michelle E McClementsNuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neurosciences and NIHR Oxford Biomedical Research Centre, University of Oxford, Oxford, United Kingdom; and Oxford University Hospitals NHS Foundation Trust, Oxford, United Kingdom.
Robert E MacLarenNuffield Laboratory of Ophthalmology, Nuffield Department of Clinical Neurosciences and NIHR Oxford Biomedical Research Centre, University of Oxford, Oxford, United Kingdom; and Oxford University Hospitals NHS Foundation Trust, Oxford, United Kingdom.
Oxford BioMedica (United Kingdom) · GB

Funding

Medical Research Council MC_PC_18059
6 · The paper itself

Abstract

Adeno-associated virus (AAV) vectors have been widely adopted for delivery of CRISPR-Cas components, especially for therapeutic gene editing. For a single vector system, both the Cas9 and guide RNA (gRNA) are encoded within a single transgene, usually from separate promoters. Careful design of this bi-cistronic construct is required due to the minimal packaging capacity of AAV. We investigated how placement of the U6 promoter expressing the gRNA on the reverse strand to SaCas9 driven by a cytomegalovirus promoter affected gene editing rates compared to placement on the forward strand. We show that orientation in the reverse direction reduces editing rates from an AAV vector due to reduced transcription of both SaCas9 and guide RNA. This effect was observed only following AAV transduction; it was not seen following plasmid transfection. These results have implications for the design of AAV-CRISPR vectors, and suggest that results from optimizing plasmid transgenes may not translate when delivered via AAV.

Indexed as

Gene EditingPromoter Regions, GeneticClustered Regularly Interspaced Short Palindromic RepeatsCRISPR-Associated Protein 9CRISPR-Cas SystemsDependovirusGenetic VectorsRNA, Guide, CRISPR-Cas SystemsStaphylococcus aureusCRISPR-Associated Protein 9RNA, Guide, CRISPR-Cas Systems

Identifiers

PMID32833533
PMCPMC7469699
OpenAlexW3080086069

What OpenQuestion holds

Textmetadata
Read underepoch 390

Registered trials

None linked

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.