Evidence map›Paper›PMID 29221488›Full record

ArticleBMC research notes2017

Divergent susceptibilities to AAV-SaCas9-gRNA vector-mediated genome-editing in a single-cell-derived cell population.

Salma G Morsy, Jason M Tonne, Yaxi Zhu, Brian Lu, Karol Budzik, James W Krempski, Sherine A Ali, Mohamed A El-Feky, Yasuhiro Ikeda

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in BMC research notes, 2017. 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
–field-weighted citation impact
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.

  1. Review
  2. Article
  3. Review
  4. Article
  5. Review
  6. Review
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

9 authors.

Salma G MorsyDepartment of Molecular Medicine, Mayo Clinic, College of Medicine, 200 First St. SW, Rochester, MN, 55905, USA.
Jason M TonneDepartment of Molecular Medicine, Mayo Clinic, College of Medicine, 200 First St. SW, Rochester, MN, 55905, USA.
Yaxi ZhuDepartment of Molecular Medicine, Mayo Clinic, College of Medicine, 200 First St. SW, Rochester, MN, 55905, USA.
Brian LuDepartment of Molecular Medicine, Mayo Clinic, College of Medicine, 200 First St. SW, Rochester, MN, 55905, USA.
Karol BudzikDepartment of Molecular Medicine, Mayo Clinic, College of Medicine, 200 First St. SW, Rochester, MN, 55905, USA.
James W KrempskiDepartment of Molecular Medicine, Mayo Clinic, College of Medicine, 200 First St. SW, Rochester, MN, 55905, USA.
Sherine A AliMicrobiology and Immunology, Faculty of Medicine, Assiut University, Assiut, Egypt.
Mohamed A El-FekyMicrobiology and Immunology, Faculty of Medicine, Assiut University, Assiut, Egypt.
Yasuhiro IkedaDepartment of Molecular Medicine, Mayo Clinic, College of Medicine, 200 First St. SW, Rochester, MN, 55905, USA. ikeda.yasuhiro@mayo.edu.

Funding

PHD TRAINING PROGRAM IN BASIC IMMUNOLOGYT32AI007425 · NIAID · MAYO CLINIC ROCHESTER · PI SHAPIRO, VIRGINIA SMITH · 1995 to 2020
$3.6M
Regulated Long-Term Cardiac BNP Expression in Hypertensive Heart DiseaseR01HL098502 · NHLBI · MAYO CLINIC ROCHESTER · PI CATALIOTTI, ALESSANDRO, IKEDA, YASUHIRO · 2010 to 2014
$1.9M
National Institutes of Health (US) HL098502NHLBI NIH HHS R01 HL098502NIAID NIH HHS T32 AI007425
6 · The paper itself

Abstract

objectiveRecombinant adeno-associated virus (AAV)-based vectors are characterized by their robust and safe transgene delivery. The CRISPR/Cas9 and guide RNA (gRNA) system present a promising genome-editing platform, and a recent development of a shorter Cas9 enzyme from Staphylococcus aureus (SaCas9) allows generation of high titer single AAV vectors which carry both saCas9- and gRNA-expression cassettes. Here, we used two AAV-SaCas9 vectors with distinct GFP-targeted gRNA sequences and determined the impact of AAV-SaCas9-gRNA vector treatment in a single cell clone carrying a GFP-expression cassette.

resultsOur results showed comparable GFP knockout efficiencies (40-50%) upon a single low-dose infection. Three consecutive transductions of 25-fold higher doses of vectors showed 80% GFP knockout efficiency. To analyze the "AAV-SaCas9-resistant cell population", we sorted the residual GFP-positive cells and assessed their permissiveness to super-infection with two AAV-Cas9-GFP vectors. We found the sorted cells were significantly more resistant to the GFP knockout mediated by the same AAV vector, but not by the other GFP-targeted AAV vector. Our data therefore demonstrate highly efficient genome-editing by the AAV-SaCas9-gRNA vector system. Differential susceptibilities of single cell-derived cells to the AAV-SaCas9-gRNA-mediated genome editing may represent a formidable barrier to achieve 100% genome editing efficiency by this vector system.

Indexed as

CRISPR-Cas SystemsDependovirusExome SequencingGene EditingGenetic VectorsRNA, Guide, CRISPR-Cas SystemsBacterial ProteinsCell LineCRISPR-Associated Protein 9CRISPR-Associated ProteinsDisease SusceptibilityEndonucleasesGreen Fluorescent ProteinsHEK293 CellsHumansStaphylococcus aureusBacterial ProteinsCas9 endonuclease Streptococcus pyogenesCRISPR-Associated Protein 9CRISPR-Associated ProteinsEndonucleasesGreen Fluorescent ProteinsRNA, Guide, CRISPR-Cas SystemsAdeno Associated viral vectorGenome editing resistanceOff-targetSaCas9 optimizationWhole exome sequencing

Identifiers

PMID29221488
PMCPMC5721606

What OpenQuestion holds

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LicenceCC BY
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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.