Characterization of Staphylococcus aureus Cas9: a smaller Cas9 for all-in-one adeno-associated virus delivery and paired nickase applications.

Abstract:

BACKGROUND:CRISPR-Cas systems have been broadly embraced as effective tools for genome engineering applications, with most studies to date utilizing the Streptococcus pyogenes Cas9. Here we characterize and manipulate the smaller, 1053 amino acid nuclease Staphylococcus aureus Cas9. RESULTS:We find that the S. aureus Cas9 recognizes an NNGRRT protospacer adjacent motif (PAM) and cleaves target DNA at high efficiency with a variety of guide RNA (gRNA) spacer lengths. When directed against genomic targets with mutually permissive NGGRRT PAMs, the S. pyogenes Cas9 and S. aureus Cas9 yield indels at comparable rates. We additionally show D10A and N580A paired nickase activity with S. aureus Cas9, and we further package it with two gRNAs in a single functional adeno-associated virus (AAV) vector. Finally, we assess comparative S. pyogenes and S. aureus Cas9 specificity using GUIDE-seq. CONCLUSION:Our results reveal an S. aureus Cas9 that is effective for a variety of genome engineering purposes, including paired nickase approaches and all-in-one delivery of Cas9 and multiple gRNA expression cassettes with AAV vectors.

journal_name

Genome Biol

journal_title

Genome biology

authors

Friedland AE,Baral R,Singhal P,Loveluck K,Shen S,Sanchez M,Marco E,Gotta GM,Maeder ML,Kennedy EM,Kornepati AV,Sousa A,Collins MA,Jayaram H,Cullen BR,Bumcrot D

doi

10.1186/s13059-015-0817-8

subject

Has Abstract

pub_date

2015-11-24 00:00:00

pages

257

eissn

1474-7596

issn

1474-760X

pii

10.1186/s13059-015-0817-8

journal_volume

16

pub_type

杂志文章
  • Is mouse embryonic stem cell technology obsolete?

    abstract::Injection of recombinant Cas9 protein and synthetic guide RNAs into mouse zygotes has been shown to facilitate gene disruption and knock-ins using the CRISPR system. These technologies may soon displace genetic modification using embryonic stem cells. ...

    journal_title:Genome biology

    pub_type: 评论,杂志文章

    doi:10.1186/s13059-015-0673-6

    authors: Skarnes WC

    更新日期:2015-05-27 00:00:00

  • Genome-wide analyses of Shavenbaby target genes reveals distinct features of enhancer organization.

    abstract:BACKGROUND:Developmental programs are implemented by regulatory interactions between Transcription Factors (TFs) and their target genes, which remain poorly understood. While recent studies have focused on regulatory cascades of TFs that govern early development, little is known about how the ultimate effectors of cell...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2013-14-8-r86

    authors: Menoret D,Santolini M,Fernandes I,Spokony R,Zanet J,Gonzalez I,Latapie Y,Ferrer P,Rouault H,White KP,Besse P,Hakim V,Aerts S,Payre F,Plaza S

    更新日期:2013-08-23 00:00:00

  • Host-associated microbiomes are predicted by immune system complexity and climate.

    abstract:BACKGROUND:Host-associated microbiomes, the microorganisms occurring inside and on host surfaces, influence evolutionary, immunological, and ecological processes. Interactions between host and microbiome affect metabolism and contribute to host adaptation to changing environments. Meta-analyses of host-associated bacte...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/s13059-019-1908-8

    authors: Woodhams DC,Bletz MC,Becker CG,Bender HA,Buitrago-Rosas D,Diebboll H,Huynh R,Kearns PJ,Kueneman J,Kurosawa E,LaBumbard BC,Lyons C,McNally K,Schliep K,Shankar N,Tokash-Peters AG,Vences M,Whetstone R

    更新日期:2020-02-03 00:00:00

  • Design and computational analysis of single-cell RNA-sequencing experiments.

    abstract::Single-cell RNA-sequencing (scRNA-seq) has emerged as a revolutionary tool that allows us to address scientific questions that eluded examination just a few years ago. With the advantages of scRNA-seq come computational challenges that are just beginning to be addressed. In this article, we highlight the computational...

    journal_title:Genome biology

    pub_type: 杂志文章,评审

    doi:10.1186/s13059-016-0927-y

    authors: Bacher R,Kendziorski C

    更新日期:2016-04-07 00:00:00

  • Histone modifications: from genome-wide maps to functional insights.

    abstract::A large number of histone modifications has been implicated in the regulation of gene expression. Together, these modifications have the potential to form a complex combinatorial regulatory code. Genome-wide mapping approaches provide new opportunities to decipher this code, but they may suffer from systematic biases....

    journal_title:Genome biology

    pub_type: 杂志文章,评审

    doi:10.1186/gb-2005-6-6-113

    authors: van Leeuwen F,van Steensel B

    更新日期:2005-01-01 00:00:00

  • Cloning and characterization of microRNAs from wheat (Triticum aestivum L.).

    abstract:BACKGROUND:MicroRNAs (miRNAs) are a class of small, non-coding regulatory RNAs that regulate gene expression by guiding target mRNA cleavage or translational inhibition. So far, identification of miRNAs has been limited to a few model plant species, such as Arabidopsis, rice and Populus, whose genomes have been sequenc...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2007-8-6-r96

    authors: Yao Y,Guo G,Ni Z,Sunkar R,Du J,Zhu JK,Sun Q

    更新日期:2007-01-01 00:00:00

  • Variation in gene duplicates with low synonymous divergence in Saccharomyces cerevisiae relative to Caenorhabditis elegans.

    abstract:BACKGROUND:The direct examination of large, unbiased samples of young gene duplicates in their early stages of evolution is crucial to understanding the origin, divergence and preservation of new genes. Furthermore, comparative analysis of multiple genomes is necessary to determine whether patterns of gene duplication ...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2009-10-7-r75

    authors: Katju V,Farslow JC,Bergthorsson U

    更新日期:2009-01-01 00:00:00

  • Species-wide distribution of highly polymorphic minisatellite markers suggests past and present genetic exchanges among house mouse subspecies.

    abstract:BACKGROUND:Four hypervariable minisatellite loci were scored on a panel of 116 individuals of various geographical origins representing a large part of the diversity present in house mouse subspecies. Internal structures of alleles were determined by minisatellite variant repeat mapping PCR to produce maps of interming...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2007-8-5-r80

    authors: Bonhomme F,Rivals E,Orth A,Grant GR,Jeffreys AJ,Bois PR

    更新日期:2007-01-01 00:00:00

  • Functional constraint and small insertions and deletions in the ENCODE regions of the human genome.

    abstract:BACKGROUND:We describe the distribution of indels in the 44 Encyclopedia of DNA Elements (ENCODE) regions (about 1% of the human genome) and evaluate the potential contributions of small insertion and deletion polymorphisms (indels) to human genetic variation. We relate indels to known genomic annotation features and m...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2007-8-9-r180

    authors: Clark TG,Andrew T,Cooper GM,Margulies EH,Mullikin JC,Balding DJ

    更新日期:2007-01-01 00:00:00

  • Personalized and graph genomes reveal missing signal in epigenomic data.

    abstract:BACKGROUND:Epigenomic studies that use next generation sequencing experiments typically rely on the alignment of reads to a reference sequence. However, because of genetic diversity and the diploid nature of the human genome, we hypothesize that using a generic reference could lead to incorrectly mapped reads and bias ...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/s13059-020-02038-8

    authors: Groza C,Kwan T,Soranzo N,Pastinen T,Bourque G

    更新日期:2020-05-25 00:00:00

  • Fine-tuning sugar content in strawberry.

    abstract::Fine-tuning quantitative traits for continuous subtle phenotypes is highly advantageous. We engineer the highly conserved upstream open reading frame (uORF) of FvebZIPs1.1 in strawberry (Fragaria vesca), using base editor A3A-PBE. Seven novel alleles are generated. Sugar content of the homozygous T1 mutant lines is 33...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/s13059-020-02146-5

    authors: Xing S,Chen K,Zhu H,Zhang R,Zhang H,Li B,Gao C

    更新日期:2020-09-03 00:00:00

  • FineMAV: prioritizing candidate genetic variants driving local adaptations in human populations.

    abstract::We present a new method, Fine-Mapping of Adaptive Variation (FineMAV), which combines population differentiation, derived allele frequency, and molecular functionality to prioritize positively selected candidate variants for functional follow-up. We calibrate and test FineMAV using eight experimentally validated "gold...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/s13059-017-1380-2

    authors: Szpak M,Mezzavilla M,Ayub Q,Chen Y,Xue Y,Tyler-Smith C

    更新日期:2018-01-17 00:00:00

  • xGDB: open-source computational infrastructure for the integrated evaluation and analysis of genome features.

    abstract::The eXtensible Genome Data Broker (xGDB) provides a software infrastructure consisting of integrated tools for the storage, display, and analysis of genome features in their genomic context. Common features include gene structure annotations, spliced alignments, mapping of repetitive sequence, and microarray probes, b...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2006-7-11-r111

    authors: Schlueter SD,Wilkerson MD,Dong Q,Brendel V

    更新日期:2006-01-01 00:00:00

  • Quantitative reconstruction of leukocyte subsets using DNA methylation.

    abstract:BACKGROUND:Cell lineage-specific DNA methylation patterns distinguish normal human leukocyte subsets and can be used to detect and quantify these subsets in peripheral blood. We have developed an approach that uses DNA methylation to simultaneously quantify multiple leukocyte subsets, enabling investigation of immune m...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2014-15-3-r50

    authors: Accomando WP,Wiencke JK,Houseman EA,Nelson HH,Kelsey KT

    更新日期:2014-03-05 00:00:00

  • SCALE: modeling allele-specific gene expression by single-cell RNA sequencing.

    abstract::Allele-specific expression is traditionally studied by bulk RNA sequencing, which measures average expression across cells. Single-cell RNA sequencing allows the comparison of expression distribution between the two alleles of a diploid organism and the characterization of allele-specific bursting. Here, we propose SC...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/s13059-017-1200-8

    authors: Jiang Y,Zhang NR,Li M

    更新日期:2017-04-26 00:00:00

  • Network security and data integrity in academia: an assessment and a proposal for large-scale archiving.

    abstract::A direct impediment to the optimal use of online databases is the increasing prevalence, severity, and toll of computer and network security incidents. Funding agencies should set up working groups that can provide essential services such as universal backup, archival storage, and mirroring of community resources, con...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2005-6-9-119

    authors: Smith A,Greenbaum D,Douglas SM,Long M,Gerstein M

    更新日期:2005-01-01 00:00:00

  • Roles of piRNAs in transposon and pseudogene regulation of germline mRNAs and lncRNAs.

    abstract::PIWI proteins, a subfamily of PAZ/PIWI Domain family RNA-binding proteins, are best known for their function in silencing transposons and germline development by partnering with small noncoding RNAs called PIWI-interacting RNAs (piRNAs). However, recent studies have revealed multifaceted roles of the PIWI-piRNA pathwa...

    journal_title:Genome biology

    pub_type: 杂志文章,评审

    doi:10.1186/s13059-020-02221-x

    authors: Wang C,Lin H

    更新日期:2021-01-08 00:00:00

  • Schmutzi: estimation of contamination and endogenous mitochondrial consensus calling for ancient DNA.

    abstract:UNLABELLED:Ancient DNA is typically highly degraded with appreciable cytosine deamination, and contamination with present-day DNA often complicates the identification of endogenous molecules. Together, these factors impede accurate assembly of the endogenous ancient mitochondrial genome. We present schmutzi, an iterati...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/s13059-015-0776-0

    authors: Renaud G,Slon V,Duggan AT,Kelso J

    更新日期:2015-10-12 00:00:00

  • Functional and regulatory profiling of energy metabolism in fission yeast.

    abstract:BACKGROUND:The control of energy metabolism is fundamental for cell growth and function and anomalies in it are implicated in complex diseases and ageing. Metabolism in yeast cells can be manipulated by supplying different carbon sources: yeast grown on glucose rapidly proliferates by fermentation, analogous to tumour ...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/s13059-016-1101-2

    authors: Malecki M,Bitton DA,Rodríguez-López M,Rallis C,Calavia NG,Smith GC,Bähler J

    更新日期:2016-11-25 00:00:00

  • Studying the microbiology of the indoor environment.

    abstract::The majority of people in the developed world spend more than 90% of their lives indoors. Here, we examine our understanding of the bacteria that co-inhabit our artificial world and how they might influence human health. ...

    journal_title:Genome biology

    pub_type: 杂志文章,评审

    doi:10.1186/gb-2013-14-2-202

    authors: Kelley ST,Gilbert JA

    更新日期:2013-02-28 00:00:00

  • Species-specific shifts in centromere sequence composition are coincident with breakpoint reuse in karyotypically divergent lineages.

    abstract:BACKGROUND:It has been hypothesized that rapid divergence in centromere sequences accompanies rapid karyotypic change during speciation. However, the reuse of breakpoints coincident with centromeres in the evolution of divergent karyotypes poses a potential paradox. In distantly related species where the same centromer...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2007-8-8-r170

    authors: Bulazel KV,Ferreri GC,Eldridge MD,O'Neill RJ

    更新日期:2007-01-01 00:00:00

  • Repression of chimeric transcripts emanating from endogenous retrotransposons by a sequence-specific transcription factor.

    abstract:BACKGROUND:Retroviral elements are pervasively transcribed and dynamically regulated during development. While multiple histone- and DNA-modifying enzymes have broadly been associated with their global silencing, little is known about how the many diverse retroviral families are each selectively recognized. RESULTS:He...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2014-15-4-r58

    authors: Mak KS,Burdach J,Norton LJ,Pearson RC,Crossley M,Funnell AP

    更新日期:2014-04-30 00:00:00

  • Genotyping structural variants in pangenome graphs using the vg toolkit.

    abstract::Structural variants (SVs) remain challenging to represent and study relative to point mutations despite their demonstrated importance. We show that variation graphs, as implemented in the vg toolkit, provide an effective means for leveraging SV catalogs for short-read SV genotyping experiments. We benchmark vg against...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/s13059-020-1941-7

    authors: Hickey G,Heller D,Monlong J,Sibbesen JA,Sirén J,Eizenga J,Dawson ET,Garrison E,Novak AM,Paten B

    更新日期:2020-02-12 00:00:00

  • Mammalian conserved ADAR targets comprise only a small fragment of the human editosome.

    abstract:BACKGROUND:ADAR proteins are among the most extensively studied RNA binding proteins. They bind to their target and deaminate specific adenosines to inosines. ADAR activity is essential, and the editing of a subset of their targets is critical for viability. Recently, a huge number of novel ADAR targets were detected b...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2014-15-1-r5

    authors: Pinto Y,Cohen HY,Levanon EY

    更新日期:2014-01-07 00:00:00

  • Ancient DNA and the rewriting of human history: be sparing with Occam's razor.

    abstract::Ancient DNA research is revealing a human history far more complex than that inferred from parsimonious models based on modern DNA. Here, we review some of the key events in the peopling of the world in the light of the findings of work on ancient DNA. ...

    journal_title:Genome biology

    pub_type: 历史文章,杂志文章,评审

    doi:10.1186/s13059-015-0866-z

    authors: Haber M,Mezzavilla M,Xue Y,Tyler-Smith C

    更新日期:2016-01-11 00:00:00

  • Broad network-based predictability of Saccharomyces cerevisiae gene loss-of-function phenotypes.

    abstract::We demonstrate that loss-of-function yeast phenotypes are predictable by guilt-by-association in functional gene networks. Testing 1,102 loss-of-function phenotypes from genome-wide assays of yeast reveals predictability of diverse phenotypes, spanning cellular morphology, growth, metabolism, and quantitative cell sha...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2007-8-12-r258

    authors: McGary KL,Lee I,Marcotte EM

    更新日期:2007-01-01 00:00:00

  • Butterfly gene flow goes berserk.

    abstract::A new study shows that genomic introgression between two Heliconius butterfly species is not solely confined to color pattern loci. ...

    journal_title:Genome biology

    pub_type: 评论,杂志文章

    doi:10.1186/s13059-016-0898-z

    authors: Ffrench-Constant RH

    更新日期:2016-02-27 00:00:00

  • A comparison of automatic cell identification methods for single-cell RNA sequencing data.

    abstract:BACKGROUND:Single-cell transcriptomics is rapidly advancing our understanding of the cellular composition of complex tissues and organisms. A major limitation in most analysis pipelines is the reliance on manual annotations to determine cell identities, which are time-consuming and irreproducible. The exponential growt...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/s13059-019-1795-z

    authors: Abdelaal T,Michielsen L,Cats D,Hoogduin D,Mei H,Reinders MJT,Mahfouz A

    更新日期:2019-09-09 00:00:00

  • Extensive transcriptomic and epigenomic remodelling occurs during Arabidopsis thaliana germination.

    abstract:BACKGROUND:Seed germination involves progression from complete metabolic dormancy to a highly active, growing seedling. Many factors regulate germination and these interact extensively, forming a complex network of inputs that control the seed-to-seedling transition. Our understanding of the direct regulation of gene e...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/s13059-017-1302-3

    authors: Narsai R,Gouil Q,Secco D,Srivastava A,Karpievitch YV,Liew LC,Lister R,Lewsey MG,Whelan J

    更新日期:2017-09-15 00:00:00

  • Transcriptome analysis of human tissues and cell lines reveals one dominant transcript per gene.

    abstract:BACKGROUND:RNA sequencing has opened new avenues for the study of transcriptome composition. Significant evidence has accumulated showing that the human transcriptome contains in excess of a hundred thousand different transcripts. However, it is still not clear to what extent this diversity prevails when considering th...

    journal_title:Genome biology

    pub_type: 杂志文章

    doi:10.1186/gb-2013-14-7-r70

    authors: Gonzàlez-Porta M,Frankish A,Rung J,Harrow J,Brazma A

    更新日期:2013-07-01 00:00:00