RNA-Guided Human Genome Engineering via Cas9
2013 · paper · Biomedicine
Adapts the bacterial CRISPR/Cas9 system to work with custom guide RNAs in human cells. It reports targeting rates of a few percent to 25% at one locus across cell types, multiplex editing, and a computed library of about 190,000 guide RNAs.
1 finding
Substance Abstract prints K562 targeting rates as "13 to 8% in K562 cells"; the Results text ("13 and 38% in K562s"), the Fig. 2 legend ("13% and 38% in K562") and the Fig. 2B panels (13.3%, 37.6%) give 13 and 38%.
- p. 1 Abstract
13 to 8% in K562 cells
- p. 2 Results, native AAVS1 locus paragraph
13 and 38% in K562s
- p. 8 Fig. 2 legend (B)
13% and 38% in K562
Judge's note: Viewed the abstract crop: the image reads "13 to 8%", so this is not an extraction artefact. Page 2 text and the page 8 legend both say 38%, and the Fig. 2B K562 panels are labelled 13.3% and 37.6%. The other two ranges in the same abstract sentence (10 to 25%, 2 to 4%) match the T1/T2 NHEJ rates, so it is the same quantity. No correction notice is among the files. The evident fix is "13 to 38%".
Second check: another model agreed
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6 minor (typos and the like; most checked by the judge only)
Wording The efficiency comparison "TALENs, while offering similar or greater efficiencies (4)" is cited to reference 4 (Jinek et al., "A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity"), whose title does not indicate a ZFN/TALEN efficiency comparison.
- p. 3 Discussion, first paragraph
TALENs, while offering similar or greater efficiencies (4)
- p. 4 Reference 4
Jinek M, et al. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial
- p. 4 Reference list, entry 4
Jinek M, et al. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity
- p. 3 Discussion
offering similar or greater efficiencies (4)
Only the authors can say what is right here.
Judge's note: Rests on outside knowledge: the content of reference 4 beyond its title (that it is an in vitro bacterial Cas9 study with no ZFN/TALEN efficiency data). The page 3 image shows "(4)" directly after the efficiency claim, and again in the next sentence on CRISPR specificity. Reference 4 on page 4 is the Jinek et al. Science 2012 paper. The paper's own efficiency comparison is in Fig. 1C. The citation may be misplaced or duplicated, but only the authors can say what was intended.
Second check: another model agreed
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Wording Fig. 2C legend describes "locations of sequencing primers (arrows) for detecting successful targeted events", whereas detection in panel D is a "PCR assay 3 days after transfection", with Sanger sequencing of the PCR amplicon in panel E.
- p. 8 Fig. 2 legend (C)
locations of sequencing primers (arrows) for detecting successful targeted events
- p. 8 Fig. 2 legend (D)
PCR assay 3 days after transfection
- p. 3 Results, HR integration paragraph
by polymerase chain reaction (PCR) (Fig. 2D and fig. S10) and Sanger sequencing (Fig. 2E)
Judge's note: The page 8 image shows a convergent arrow pair in panel C, one in the genome and one in the donor, consistent with primers for a junction PCR. The legend for D says PCR assay, and E says Sanger sequencing of the PCR amplicon. The same primers could plausibly serve for both PCR and sequencing, so either wording could be legitimate; this is a minor wording inconsistency.
Second check: another model agreed
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Typo FACS is expanded as "flow-activated cell sorting (FACS)" rather than fluorescence-activated cell sorting.
- p. 2 Results, GFP reporter assay paragraph
cells by flow-activated cell sorting (FACS)
Only the authors can say what is right here.
Judge's note: Rests on outside knowledge: the standard expansion of FACS is "fluorescence-activated cell sorting". The page 2 image does read "flow-activated cell sorting (FACS)". The paper gives no other expansion to check against, so this is a wording query for the authors.
Second check: none (judge only)
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Typo Inconsistent pointer form: "(figs. S1A and fig. S3)".
- p. 3 Discussion
(figs. S1A and fig. S3)
Judge's note: The page 3 image reads "(figs. S1A and fig. S3) (4, 5)". The plural "figs." is followed by a repeated singular "fig.". Cosmetic only; the meaning is unchanged.
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Typo Fig. 1 legend says "Binding sites for the two halves of the TALEN are underlined." but in Fig. 1B the two half-sites are marked by bars above the sequence, under the label "AAVS1-TALENs target site".
- p. 7 Fig. 1 legend (B) and Fig. 1B
Binding sites for the two halves of the TALEN are underlined.
Judge's note: On the page 7 image, the two purple bars sit above the sequence line, directly under the "AAVS1-TALENs target site" label. Below the sequence are only the T1/T2 Target labels. The marks are overlines rather than underlines. This is a loose description and the reader can still identify the sites; the resolution of the page image is limited.
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Typo Subject–verb disagreement in the Fig. 2F legend: "Microscope images of two representative GFP+ clones is shown."
- p. 8 Fig. 2 legend (F)
Microscope images of two representative GFP+ clones is shown.
Judge's note: The page 8 image reads exactly as quoted. The plural subject "images" takes the singular verb "is"; it should be "are shown". Cosmetic.
Second check: none (judge only)