Genome editing has attracted wide interest for the generation of cellular models of disease using human pluripotent stem cells and other cell types. CRISPR-Cas systems and TALENs can target desired genomic sites with high efficiency in human cells, but recent publications have led to concern about the extent to which these tools may cause off-target mutagenic effects that could potentially confound disease-modeling studies. Using CRISPR-Cas9 and TALEN targeted human pluripotent stem cell clones, we performed whole-genome sequencing at high coverage in order to assess the degree of mutagenesis across the entire genome. In both types of clones, we found that off-target mutations attributable to the nucleases were very rare. From this analysis, we suggest that, although some cell types may be at risk for off-target mutations, the incidence of such effects in human pluripotent stem cells may be sufficiently low and thus not a significant concern for disease modeling and other applications.Stem Cell and Regenerative Biolog

Brand, Harrison Andrew

Collins, Ryan

Cowan, Chad A.

Ding, Qiurong

Erdin, Serkan

Gosis, Bridget S.

Musunuru, Kiran

Ragavendran, Ashok

Talkowski, Michael Edward

Veres, Adrian

Cell Stem Cell

SummaryGenome editing has attracted wide interest for the generation of cellular models of disease using human pluripotent stem cells and other cell types. CRISPR-Cas systems and TALENs can target desired genomic sites with high efficiency in human cells, but recent publications have led to concern about the extent to which these tools may cause off-target mutagenic effects that could potentially confound disease-modeling studies. Using CRISPR-Cas9 and TALEN targeted human pluripotent stem cell clones, we performed whole-genome sequencing at high coverage in order to assess the degree of mutagenesis across the entire genome. In both types of clones, we found that off-target mutations attributable to the nucleases were very rare. From this analysis, we suggest that, although some cell types may be at risk for off-target mutations, the incidence of such effects in human pluripotent stem cells may be sufficiently low and thus not a significant concern for disease modeling and other applications

Brand, Harrison

Talkowski, Michael E.

Elsevier - Publisher Connector 

Cell Stem Cell
Brief ReportLow Incidence of Off-Target Mutations in Individual
CRISPR-Cas9 and TALEN Targeted Human Stem Cell
Clones Detected by Whole-Genome Sequencing
Adrian Veres,1,2 Bridget S. Gosis,1 Qiurong Ding,1 Ryan Collins,3 Ashok Ragavendran,3 Harrison Brand,3 Serkan Erdin,3
Chad A. Cowan,1,2,5 Michael E. Talkowski,3,4,5 and Kiran Musunuru1,2,5,6,*
1Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, Cambridge, MA 02138, USA
2Department of Medicine, Harvard Medical School, Boston, MA 02115, USA
3Molecular Neurogenetics Unit, Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research,
Massachusetts General Hospital, Boston, MA 02114, USA
4Department of Neurology, Harvard Medical School, Boston, MA 02115, USA
5Broad Institute, Cambridge, MA 02142, USA
6Division of Cardiovascular Medicine, Brigham and Women’s Hospital, Boston, MA 02115, USA
*Correspondence: kiranmusunuru@gmail.com
http://dx.doi.org/10.1016/j.stem.2014.04.020SUMMARY
Genome editing has attracted wide interest for the
generation of cellular models of disease using human
pluripotent stem cells and other cell types. CRISPR-
CassystemsandTALENscan targetdesiredgenomic
sites with high efficiency in human cells, but recent
publications have led to concern about the extent
to which these tools may cause off-target mutagenic
effects that could potentially confound disease-
modeling studies. Using CRISPR-Cas9 and TALEN
targeted human pluripotent stem cell clones, we per-
formed whole-genome sequencing at high coverage
in order to assess the degree of mutagenesis across
the entire genome. In both types of clones, we found
that off-targetmutationsattributable to thenucleases
were very rare. From this analysis, we suggest that,
although some cell types may be at risk for off-target
mutations, the incidence of such effects in human
pluripotent stem cells may be sufficiently low and
thus not a significant concern for disease modeling
and other applications.Clustered regularly interspaced short palindromic repea
(CRISPR)-associated (Cas) systems and transcription acti-t
vator-like effector nucleases (TALENs) are recently developed
genome-editing tools that target desired genomic sites in
mammalian cells (Miller et al., 2011; Hockemeyer et al., 2011;
Cong et al., 2013; Mali et al., 2013b; Cho et al., 2013; Jinek
et al., 2013). The most commonly employed CRISPR-Cas sys-
tem, derived fromStreptococcus pyogenes, uses Cas9 nuclease
that complexes with a guide RNA that hybridizes a 20 nt DNA
sequence (protospacer) immediately preceding an NGG motif
(protospacer-associated motif or PAM), resulting in a double-
strand break (DSB) 3 bp upstream of the NGG (Jinek et al.,
2012). TALENs bind as a pair on sequences surrounding a
genomic site, positioning a dimer of FokI nuclease domains inorder to generate a DSB at the site. The introduction of a DSB
at a specified genomic site allows for modification of the site
via either nonhomologous end-joining (NHEJ), which typically in-
troduces an insertion or deletion (indel), or homology-directed
repair, which can be exploited to knock in a point mutation or
insert a desired sequence at the site.
One important application of genome-editing technology is
disease modeling (Musunuru, 2013). The ability to generate
isogenicwild-type andmutant clones for phenotypic comparison
would enable rigorous functional genetic studies. However, both
CRISPR-Cas9 and TALENs have been demonstrated to produce
off-target effects; i.e., mutagenesis at sites in the genome other
than the desired on-target site (Hockemeyer et al., 2011; Musso-
lino et al., 2011; Fu et al., 2013; Hsu et al., 2013;Mali et al., 2013a;
Pattanayak et al., 2013; Cradick et al., 2013; Cho et al., 2014).
These studies have largely focused on sites with high sequence
similarity to the on-target site and have documented mutagen-
esis rates as high as 77% for CRISPR-Cas9 and 1% for TALENs
at individual off-target sites. Relatively unexplored is whether
CRISPR-Cas9 or TALENs produce off-target effects at sites
with low sequence similarity to the on-target site. Although the
nucleasesmight have poor affinity and a low probability of gener-
ating a mutation at any given single site in the genome, they may
nonetheless generate a sizeable number of nonspecific muta-
tions across the billions of base pairs of the genome in any single
cell. This would significantly confound the validity of disease-
modeling studies that rely upon genome-edited clones.
To date, most studies of nuclease off-target effects have been
performed in aggregated pools of transformed or immortalized
cultured human cells such as human embryonic kidney 293T
and K562 cells that are not well suited for disease modeling.
Therefore, we decided to study nuclease off-target effects
generated in a ‘‘real-world’’ application of genome editing
centered on human pluripotent stem cell (hPSC) clones being
actively used for biological studies (e.g., Ding et al., 2013a).
We assessed the degree of genome-wide off-target muta-
genesis in hPSC clones targeted with either CRISPR-Cas9 or
TALENs. We performed whole-genome sequencing at high
coverage (603 target coverage) of ten cell lines, including nineCell Stem Cell 15, 27–30, July 3, 2014 ª2014 Elsevier Inc. 27
AB
(legend on next page)
Cell Stem Cell
Whole-Genome Sequencing of Genome-Edited Clones
28 Cell Stem Cell 15, 27–30, July 3, 2014 ª2014 Elsevier Inc.
Table 1. Numbers of Unique On-Target and Candidate Off-Target
Indels, SVs, and SNVs in TALEN and CRISPR-Cas9 Targeted
Clones
SORT1
TALENs
SORT1
CRISPR-
Cas9
LINC00116
CRISPR-
Cas9
Clones A B C D E F G H I
On-target indels — 2 2 — 1a 1a — 1 1a
On-target SVs — — — — — — — 1b —
Likely off-target indel — 1 — — — — — — —
Other candidate off-
target indels
2 1 2 4 4 2 3 5 4
Candidate off-
target SVs
— — — — — 1 — — —
SNVs 64 115 142 55 94 74 111 127 112
See also Figure S1 and Tables S1–S3.
aHomozygous for indels.
b261 bp duplicated insertion.
Cell Stem Cell
Whole-Genome Sequencing of Genome-Edited Clonesclones we had previously generated with genome editing (Ding
et al., 2013a, 2013b) (Figure 1): the human embryonic stem cell
line HUES 9: three HUES 9 clones exposed to TALENs targeting
the SORT1 gene with one clone remaining wild-type in both
alleles (clone A) and two clones bearing indels in both SORT1
alleles (clones B and C), three HUES 9 clones exposed to
CRISPR-Cas9 targeting the same site in the SORT1 gene with
one wild-type clone (clone D) and two clones bearing indels in
both SORT1 alleles (clones E and F), and three HUES 9 clones
exposed to CRISPR-Cas9 targeting the LINC00116 gene with
one wild-type clone (clone G) and two clones bearing indels in
both LINC00116 alleles (clones H and I). All of the HUES 9 clones
were derived from the same stock of parental HUES 9 cells.
Notably, we had found the targeting efficiency of the SORT1
TALENs to be 11%, in contrast to CRISPR-Cas9 for SORT1,
which was 76%, The targeting efficiency of CRISPR-Cas9 for
LINC00116 was 57% (Ding et al., 2013b).
Upon obtaining the whole-genome sequencing data, we as-
sessed the clones for small indels, single-nucleotide variants
(SNVs), and structural variants (SVs), which include chromo-
somal inversions, rearrangements, duplications, and deletions
(Supplemental Experimental Procedures available online). We
largely focused on the identification of small indels and SVs
because they comprise virtually all of the mutations introduced
by NHEJ. After filtering for the small indels most likely to be
true positives and to be potential off-target mutations (rather
than mutations that arose in the parental cell pool) and confirma-
tion with Sanger sequencing, we identified a total of 28 such
indels across the nine experimental clones in comparison to
the parental HUES 9 cells as the reference. Notably, all of the
previously known on-target indels (seven in total) were correctly
identified by the whole-genome sequencing and filtering (TablesFigure 1. On- and Off-Target Mutations
(A) HUES 9 clones targeted in the SORT1 gene with TALENs or CRISPR-Cas9.
(B) HUES 9 clones targeted in the LINC00116 genewith CRISPR-Cas9. For TALEN
sequences. For CRISPR-Cas9 targeted clones, the boxes indicate the 20 bp se
deletions and insertions in the two alleles of each clone are indicated. For the off
indicated in bold, and the deletion in one allele of the clone is indicated.1 and S1). One of the 28 off-target indels was a frameshift in the
coding sequence of ZDHHC11 (in clone I). None of the other in-
dels lay in either the coding sequence of a gene or the expressed
sequence of an annotated noncoding RNA.
None of the indels in CRISPR-Cas9 clones were within 100 nt
of a potential off-target site as predicted by sequence similar-
ity—up to six mismatches—with the on-target site, and none
lay near sequences that matched the on-target sites better
than would be expected by chance (Figure S1). Moreover,
none of the indels lay within 100 nt of a sequence perfectly
matching the last 10 nt of the protospacer with an adjacent
PAM site (NGG as well as NAG, which has also been shown to
be tolerated) (Hsu et al., 2013; Pattanayak et al., 2013). Further-
more, we paid special attention to the indels that lay within 5 bp
upstream of a potential PAM site (Table S1), where CRISPR-
Cas9-mediated DSBs would be expected to occur. Although
the majority of indels had a potential PAM site, none of the adja-
cent sequences matched the on-target site better than would be
expected by chance (Figure S1).
Oneof the indels inaTALENclonewas locatedbetween twopo-
tential off-target binding sites as predicted by sequence similarity
with the on-target sites—one with three mismatches and another
with four mismatches—with the binding sites being 17 bp apart,
within the optimal range for generating a DSB with TALENs of
this type (Ding et al., 2013a) (Figure 1A). None of the other TALEN
clone indels were optimally positioned near a pair of degenerate
TALEN binding sites (up to five mismatches with the on-target
site), and none lay near sequences that matched the on-target
sites better than would be expected by chance (Figure S1).
None of the SVs and SNVs that passed our filtering criteria in
CRISPR-Cas9 clones was within 100 nt of a predicted off-target
site. None of the variants in TALEN clones were optimally
positioned near a pair of degenerate TALEN binding sites. We
detected 894 unique SNVs across the nine clones (average of
100 per clone) in comparison to the parental HUES 9 cell line
(Table 1). The SV analysis revealed two structural variants unique
to an individual clone: a 5.5 kb deletion on chromosome 6 in
clone F and a 261 bp segment of chromosome 4 inserted within
the LINC00116 CRISPR-Cas9 on-target site on chromosome 2
in clone H (Tables 1 and S2). Sanger sequencing confirmed
that both alleles of the chromosome 4 region were intact in
clone H, signifying a duplicated insertion into the chromosome
2 on-target site rather than a balanced translocation. We specu-
late that, because of microhomology, the chromosome 4 region
was used as a repair template for a DSB at the on-target site.
Just one of the detected variants—a TALEN clone indel—
seems certain to be a nuclease-mediated off-target effect. It is
probable that some, if not all, of the other indels and SVs reflect
clonal heterogeneity within the original stock of HUES 9 cells.
Previous studies have documented mutagenesis occurring dur-
ing the derivation and expansion of hPSCs (Hussein et al., 2011;
Gore et al., 2011; Howden et al., 2011; Yusa et al., 2011).targeted clones, the boxes indicate the TALEN on-target and off-target binding
quence matching the protospacer and the 3 bp PAM. For the on-target sites,
-target site, the mismatches with the TALEN on-target binding sequences are
Cell Stem Cell 15, 27–30, July 3, 2014 ª2014 Elsevier Inc. 29
Cell Stem Cell
Whole-Genome Sequencing of Genome-Edited ClonesFurthermore, each clone harbored a sizable number of unique
SNVs, which would not be predicted to result from NHEJ. None-
theless, with a maximum of just two to five confirmed events in
each individual clone, our results suggest that nuclease-medi-
ated off-target effects of CRISPR-Cas9 and TALENs do not
intrinsically cause a large degree of indiscriminate nonspecific
mutagenesis across the genome.
We note the limitations of this study. Even with whole-genome
sequencing at high coverage, it is likely that some variants in the
clones were not detected given the limitations of short-read
sequencing. The small number of sequenced clones targeted
at just two loci prevents generalization to all hPSC clones tar-
geted with any CRISPR-Cas9 or TALENs of any configuration
by any methodology. Furthermore, our results are not relevant
to therapeutic applications targeting up to millions of cells at a
time, where rare events may have deleterious consequences.
We do note that clonal heterogeneity may represent a more
serious obstacle to the generation of truly isogenic cell lines
than nuclease-mediated off-target effects, given that each of
our clones harbored a very small number of unique indels and
SVs (two to five) in comparison to a relatively larger number of
unique SNVs (average of 100) that likely arose spontaneously
in culture. This suggests that even if one had a genome-editing
tool in hand with perfect specificity, targeted clones would still
be likely to harbor some differences elsewhere in the genome.
Rigorous studies will require whole-genome sequencing of the
clones used for experiments to fully characterize their mutational
profiles, or they will need to include multiple clones for each
experimental condition to ensure that potential confounding by
any single mutation in a clone is minimized.
ACCESSION NUMBERS
The NCBI Sequence Read Archive accession number for the whole-genome
sequencing data reported in this paper is SRP039576.
SUPPLEMENTAL INFORMATION
Supplemental Information contains Supplemental Experimental Procedures,
one figure, and three tables and can be found with this article online at
http://dx.doi.org/10.1016/j.stem.2014.04.020.
ACKNOWLEDGMENTS
This work was supported in part by the Harvard Presidential Scholars Fund of
the Harvard Medical School MD/PhD Program (A.V.); grants R00-HL098364
(K.M.), R01-HL118744 (K.M.), R01-DK097768 (K.M. and C.A.C.), U01-
HL100408 (C.A.C.), and R00-MH095867 (M.E.T.) from the National Institutes
of Health (NIH); the Harvard Stem Cell Institute (K.M.); and Harvard University
(Q.D. and K.M.). We thank Vamsee Pillalamarri, Carrie Hanscom, and the staff
of the MGH Genomics and Technology Core and NextGen Sequencing Core
for technical assistance.
Received: January 11, 2014
Revised: March 13, 2014
Accepted: April 24, 2014
Published: July 3, 2014
REFERENCES
Cho, S.W., Kim, S., Kim, J.M., and Kim, J.S. (2013). Targeted genome
engineering in human cells with the Cas9 RNA-guided endonuclease. Nat.
Biotechnol. 31, 230–232.30 Cell Stem Cell 15, 27–30, July 3, 2014 ª2014 Elsevier Inc.Cho, S.W., Kim, S., Kim, Y., Kweon, J., Kim, H.S., Bae, S., and Kim, J.S. (2014).
Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucle-
ases and nickases. Genome Res. 24, 132–141.
Cong, L., Ran, F.A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P.D., Wu, X.,
Jiang, W., Marraffini, L.A., and Zhang, F. (2013). Multiplex genome engineering
using CRISPR/Cas systems. Science 339, 819–823.
Cradick, T.J., Fine, E.J., Antico, C.J., and Bao, G. (2013). CRISPR/Cas9 sys-
tems targeting b-globin and CCR5 genes have substantial off-target activity.
Nucleic Acids Res. 41, 9584–9592.
Ding, Q., Lee, Y.K., Schaefer, E.A., Peters, D.T., Veres, A., Kim, K.,
Kuperwasser, N., Motola, D.L., Meissner, T.B., Hendriks, W.T., et al. (2013a).
A TALEN genome-editing system for generating human stem cell-based dis-
ease models. Cell Stem Cell 12, 238–251.
Ding, Q., Regan, S.N., Xia, Y., Oostrom, L.A., Cowan, C.A., and Musunuru, K.
(2013b). Enhanced efficiency of human pluripotent stem cell genome editing
through replacing TALENs with CRISPRs. Cell Stem Cell 12, 393–394.
Fu, Y., Foden, J.A., Khayter, C., Maeder, M.L., Reyon, D., Joung, J.K., and
Sander, J.D. (2013). High-frequency off-target mutagenesis induced by
CRISPR-Cas nucleases in human cells. Nat. Biotechnol. 31, 822–826.
Gore, A., Li, Z., Fung, H.L., Young, J.E., Agarwal, S., Antosiewicz-Bourget, J.,
Canto, I., Giorgetti, A., Israel, M.A., Kiskinis, E., et al. (2011). Somatic coding
mutations in human induced pluripotent stem cells. Nature 471, 63–67.
Hockemeyer, D., Wang, H., Kiani, S., Lai, C.S., Gao, Q., Cassady, J.P., Cost,
G.J., Zhang, L., Santiago, Y., Miller, J.C., et al. (2011). Genetic engineering of
human pluripotent cells using TALE nucleases. Nat. Biotechnol. 29, 731–734.
Howden, S.E., Gore, A., Li, Z., Fung, H.L., Nisler, B.S., Nie, J., Chen, G.,
McIntosh, B.E., Gulbranson, D.R., Diol, N.R., et al. (2011). Genetic correction
and analysis of induced pluripotent stem cells from a patient with gyrate atro-
phy. Proc. Natl. Acad. Sci. USA 108, 6537–6542.
Hsu, P.D., Scott, D.A., Weinstein, J.A., Ran, F.A., Konermann, S., Agarwala, V.,
Li, Y., Fine, E.J., Wu, X., Shalem, O., et al. (2013). DNA targeting specificity of
RNA-guided Cas9 nucleases. Nat. Biotechnol. 31, 827–832.
Hussein, S.M., Batada, N.N., Vuoristo, S., Ching, R.W., Autio, R., Na¨rva¨, E., Ng,
S., Sourour, M., Ha¨ma¨la¨inen, R., Olsson, C., et al. (2011). Copy number varia-
tion and selection during reprogramming to pluripotency. Nature 471, 58–62.
Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J.A., and Charpentier,
E. (2012). A programmable dual-RNA-guided DNA endonuclease in adaptive
bacterial immunity. Science 337, 816–821.
Jinek, M., East, A., Cheng, A., Lin, S., Ma, E., and Doudna, J. (2013). RNA-pro-
grammed genome editing in human cells. Elife 2, e00471.
Mali, P., Aach, J., Stranges, P.B., Esvelt, K.M., Moosburner, M., Kosuri, S.,
Yang, L., and Church, G.M. (2013a). CAS9 transcriptional activators for target
specificity screening and paired nickases for cooperative genome engineer-
ing. Nat. Biotechnol. 31, 833–838.
Mali, P., Yang, L., Esvelt, K.M., Aach, J., Guell, M., DiCarlo, J.E., Norville, J.E.,
and Church, G.M. (2013b). RNA-guided human genome engineering via Cas9.
Science 339, 823–826.
Miller, J.C., Tan, S., Qiao, G., Barlow, K.A., Wang, J., Xia, D.F., Meng, X.,
Paschon, D.E., Leung, E., Hinkley, S.J., et al. (2011). A TALE nuclease architec-
ture for efficient genome editing. Nat. Biotechnol. 29, 143–148.
Mussolino, C., Morbitzer, R., Lu¨tge, F., Dannemann, N., Lahaye, T., and
Cathomen, T. (2011). A novel TALE nuclease scaffold enables high genome
editing activity in combination with low toxicity. Nucleic Acids Res. 39,
9283–9293.
Musunuru, K. (2013). Genome editing of human pluripotent stem cells to
generate human cellular disease models. Dis. Model. Mech. 6, 896–904.
Pattanayak, V., Lin, S., Guilinger, J.P., Ma, E., Doudna, J.A., and Liu, D.R.
(2013). High-throughput profiling of off-target DNA cleavage reveals RNA-pro-
grammed Cas9 nuclease specificity. Nat. Biotechnol. 31, 839–843.
Yusa, K., Rashid, S.T., Strick-Marchand, H., Varela, I., Liu, P.Q., Paschon,
D.E., Miranda, E., Ordo´n˜ez, A., Hannan, N.R., Rouhani, F.J., et al. (2011).
Targeted gene correction of a1-antitrypsin deficiency in induced pluripotent
stem cells. Nature 478, 391–394.


Low Incidence of Off-Target Mutations in Individual CRISPR-Cas9 and TALEN Targeted Human Stem Cell Clones Detected by Whole-Genome Sequencing 

Harvard University - DASH 

Low Incidence of Off-Target Mutations in Individual CRISPR-Cas9 and TALEN Targeted Human Stem Cell Clones Detected by Whole-Genome Sequencing

Cowan, Chad A.

Cell Stem Cell
ErratumLow Incidence of Off-Target Mutations in Individual
CRISPR-Cas9 and TALEN Targeted Human Stem Cell
Clones Detected by Whole-Genome Sequencing
Adrian Veres, Bridget S. Gosis, Qiurong Ding, Ryan Collins, Ashok Ragavendran, Harrison Brand, Serkan Erdin,
Chad A. Cowan, Michael E. Talkowski, and Kiran Musunuru*
*Correspondence: kiranmusunuru@gmail.com
http://dx.doi.org/10.1016/j.stem.2014.07.009
(Cell Stem Cell 15, 27–30; July 3, 2014)
In the original version of this paper, we unfortunately neglected to include the author Chad A. Cowan, who is affiliated with the Depart-
ment of StemCell and Regenerative Biology at Harvard University and the Harvard StemCell Institute, the Department of Medicine at
Harvard Medical School, and the Broad Institute.
The correct author list appears above. The online version of the paper has also been corrected to reflect this change.
Additionally, a corrected Acknowledgments section now appears both in the online version of the paper and as follows:
This work was supported in part by the Harvard Presidential Scholars Fund of the Harvard Medical School MD/PhD Program (A.V.);
grants R00-HL098364 (K.M.), R01-HL118744 (K.M.), R01-DK097768 (K.M. and C.A.C.), U01-HL100408 (C.A.C.), and R00-
MH095867 (M.E.T.) from the National Institutes of Health (NIH); the Harvard Stem Cell Institute (K.M.); and Harvard University
(Q.D. and K.M.). We thank Vamsee Pillalamarri, Carrie Hanscom, and the staff of the MGH Genomics and Technology Core and
NextGen Sequencing Core for technical assistance.254 Cell Stem Cell 15, 254, August 7, 2014 ª2014 Elsevier Inc.


file:///data/remote/core/dit/data/elsevier/pdf/07b/aHR0cDovL2FwaS5lbHNldmllci5jb20vY29udGVudC9hcnRpY2xlL3BpaS9zMTkzNDU5MDkxNDAwMTg2Ng==.pdf

Low Incidence of Off-Target Mutations in Individual CRISPR-Cas9 and TALEN Targeted Human Stem Cell Clones Detected by Whole-Genome Sequencing

Abstract

Similar works

Full text

Available Versions

Elsevier - Publisher Connector

Harvard University - DASH

Elsevier - Publisher Connector

Elsevier - Publisher Connector

Elsevier - Publisher Connector