CRISPR-Cas9-guided oncogenic chromosomal translocations with conditional fusion protein expression in human mesenchymal cells.

Abstract:

:Gene editing techniques have been extensively used to attempt to model recurrent genomic rearrangements found in tumor cells. These methods involve the induction of double-strand breaks at endogenous loci followed by the identification of breakpoint junctions within a population, which typically arise by nonhomologous end joining. The low frequency of these events, however, has hindered the cloning of cells with the desired rearrangement before oncogenic transformation. Here we present a strategy combining CRISPR-Cas9 technology and homology-directed repair to allow for the selection of human mesenchymal stem cells harboring the oncogenic translocation EWSR1-WT1 found in the aggressive desmoplastic small round cell tumor. The expression of the fusion transcript is under the control of the endogenous EWSR1 promoter and, importantly, can be conditionally expressed using Cre recombinase. This method is easily adapted to generate any cancer-relevant rearrangement.

authors

Vanoli F,Tomishima M,Feng W,Lamribet K,Babin L,Brunet E,Jasin M

doi

10.1073/pnas.1700622114

subject

Has Abstract

pub_date

2017-04-04 00:00:00

pages

3696-3701

issue

14

eissn

0027-8424

issn

1091-6490

pii

1700622114

journal_volume

114

pub_type

杂志文章
  • RNA-dependent RNA polymerase consensus sequence of the L-A double-stranded RNA virus: definition of essential domains.

    abstract::The L-A double-stranded RNA virus of Saccharomyces cerevisiae makes a gag-pol fusion protein by a -1 ribosomal frameshift. The pol amino acid sequence includes consensus patterns typical of the RNA-dependent RNA polymerases (EC 2.7.7.48) of (+) strand and double-stranded RNA viruses of animals and plants. We have carr...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.89.6.2185

    authors: Ribas JC,Wickner RB

    更新日期:1992-03-15 00:00:00

  • Diazonamide toxins reveal an unexpected function for ornithine delta-amino transferase in mitotic cell division.

    abstract::We have studied a naturally occurring small-molecule antimitotic called diazonamide A. Diazonamide A is highly effective at blocking spindle assembly in mammalian cell culture and does so through a unique mechanism. A biotinylated form of diazonamide A affinity purifies ornithine delta-amino transferase (OAT), a mitoc...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0610832104

    authors: Wang G,Shang L,Burgett AW,Harran PG,Wang X

    更新日期:2007-02-13 00:00:00

  • Differential requirements of MHC and of DCs for endogenous proliferation of different T-cell subsets in vivo.

    abstract::T cells transferred into severe lymphopenic hosts undergo rapid proliferation known as "endogenous proliferation" that are distinct from conventional homeostatic proliferation. Unlike homeostatic proliferation, cytokines, such as IL-7 are dispensable, yet TCR:MHC interaction is essential for this process to occur. How...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0909954106

    authors: Do JS,Min B

    更新日期:2009-12-01 00:00:00

  • Multiple ligands for cytoadherence can be present simultaneously on the surface of Plasmodium falciparum-infected erythrocytes.

    abstract::A major virulence factor of Plasmodium falciparum is the adherence of parasitized erythrocytes to the wall of postcapillary venules via a specific interaction between parasite-derived erythrocyte surface ligands and receptors on endothelial cells. To study this phenomenon in vitro, we selected a parasite population th...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.91.23.10805

    authors: Chaiyaroj SC,Coppel RL,Novakovic S,Brown GV

    更新日期:1994-11-08 00:00:00

  • Truncated erythropoietin receptor causes dominantly inherited benign human erythrocytosis.

    abstract::Erythropoietin regulates the proliferation and differentiation of erythroid precursor cells. Its effect is mediated by the erythropoietin receptor (EPOR), a member of a large family of cytokine receptors. The EPOR gene has recently been cloned, sequenced, and characterized. As shown experimentally, its intracellular C...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.90.10.4495

    authors: de la Chapelle A,Träskelin AL,Juvonen E

    更新日期:1993-05-15 00:00:00

  • Regulation of antibody heterogeneity by suppressor T cells: diminishing suppressor T cell activity increases the number of dinitrophenyl clones in mice immunized with dinitrophenyl-poly(Glu,Lys,Phe) or dinitrophenyl-poly(Glu,Lys,Ala).

    abstract::The anti-hapten responses to the dinitrophenyl (Dnp) conjugate of the random linear terpolymers poly(L-Glu,LLys,LPhe), GLphi, or poly(LGlu,LLys,LAla), GLA, are of highly restricted heterogeneity. Thus, individual mice of most responder strains express an average of two to four anti-Dnp clones to Dnp-GLphi or Dnp-GLA, ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.75.6.2914

    authors: Kipps TJ,Benacerraf B,Dorf ME

    更新日期:1978-06-01 00:00:00

  • Uncoupling of the DNA topoisomerase and replication activities of an initiator protein.

    abstract::The replication initiator proteins encoded by the pT181 and related plasmids have sequence-specific DNA binding and topoisomerase activities. These proteins create a site-specific nick in one strand of the DNA at the origin of replication that serves as a primer for the initiation of replication. To define the regions...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.89.7.3083

    authors: Dempsey LA,Birch P,Khan SA

    更新日期:1992-04-01 00:00:00

  • Immortalized keratinocyte lines derived from human embryonic stem cells.

    abstract::Cells of the human embryonic stem (hES) cell line H9, when cultured in the form of embryoid bodies, give rise to cells with markers of the keratinocyte of stratified squamous epithelia. Keratinocytes also form in nodules produced in scid mice by injected H9 cells; the hES-derived keratinocytes could be recovered in cu...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0510953103

    authors: Iuchi S,Dabelsteen S,Easley K,Rheinwald JG,Green H

    更新日期:2006-02-07 00:00:00

  • Pleiotropic effects for Parkin and LRRK2 in leprosy type-1 reactions and Parkinson's disease.

    abstract::Type-1 reactions (T1R) are pathological inflammatory episodes and main contributors to nerve damage in leprosy. Here, we evaluate the genewise enrichment of rare protein-altering variants in 7 genes where common variants were previously associated with T1R. We selected 474 Vietnamese leprosy patients of which 237 were...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 临床试验,杂志文章,多中心研究

    doi:10.1073/pnas.1901805116

    authors: Fava VM,Xu YZ,Lettre G,Van Thuc N,Orlova M,Thai VH,Tao S,Croteau N,Eldeeb MA,MacDougall EJ,Cambri G,Lahiri R,Adams L,Fon EA,Trempe JF,Cobat A,Alcaïs A,Abel L,Schurr E

    更新日期:2019-07-30 00:00:00

  • Social evolution in multispecies biofilms.

    abstract::Microbial ecology is revealing the vast diversity of strains and species that coexist in many environments, ranging from free-living communities to the symbionts that compose the human microbiome. In parallel, there is growing evidence of the importance of cooperative phenotypes for the growth and behavior of microbia...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1100292108

    authors: Mitri S,Xavier JB,Foster KR

    更新日期:2011-06-28 00:00:00

  • Alkylating agent resistance: in vitro studies with human cell lines.

    abstract::Development of in vitro resistance to HN2 (also called mustargen or mechlorethamine hydrochloride), N,N'-bis(2-chloroethyl)-N-nitrosourea (BCNU), and cisplatin [cis-diamminedichloroplatinum(II)] was achieved in two human cell lines, the Raji/Burkitt lymphoma and a squamous cell carcinoma of the tongue. A 10- to 20-fol...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.82.7.2158

    authors: Frei E 3rd,Cucchi CA,Rosowsky A,Tantravahi R,Bernal S,Ervin TJ,Ruprecht RM,Haseltine WA

    更新日期:1985-04-01 00:00:00

  • Stratification of reactivity determines nitrate removal in groundwater.

    abstract::Biogeochemical reactions occur unevenly in space and time, but this heterogeneity is often simplified as a linear average due to sparse data, especially in subsurface environments where access is limited. For example, little is known about the spatial variability of groundwater denitrification, an important process in...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1816892116

    authors: Kolbe T,de Dreuzy JR,Abbott BW,Aquilina L,Babey T,Green CT,Fleckenstein JH,Labasque T,Laverman AM,Marçais J,Peiffer S,Thomas Z,Pinay G

    更新日期:2019-02-12 00:00:00

  • Involvement of estrogen receptor beta in terminal differentiation of mammary gland epithelium.

    abstract::The mammary glands of prepubertal estrogen receptor (ER)beta-- mice are morphologically indistinguishable from those of WT littermates. It appears that, although ERbeta is expressed in the mouse mammary gland, it is not involved in ductal growth of the gland. In this study, we examined the possibility that ERbeta has ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.192561299

    authors: Förster C,Mäkela S,Wärri A,Kietz S,Becker D,Hultenby K,Warner M,Gustafsson JA

    更新日期:2002-11-26 00:00:00

  • The gene for an inherited form of deafness maps to chromosome 5q31.

    abstract::Primary--i.e., nonsyndromal-postlingual deafness is inherited as an autosomal dominant phenotype in a large kindred in Costa Rica. Genetically susceptible individuals begin to lose hearing at low frequencies at about age 10 years, after language and speaking are learned. Deafness inevitably progresses by age 30 years ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.89.11.5181

    authors: Leon PE,Raventos H,Lynch E,Morrow J,King MC

    更新日期:1992-06-01 00:00:00

  • Toward development of a screen to identify randomly encoded, foldable sequences.

    abstract::The ability to identify sequences in a randomly encoded polypeptide library that are capable of acquiring unique and stably folded structures would be valuable in the examination of protein-folding issues. The quality control system of the yeast secretory pathway prevents the release of incompletely folded polypeptide...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.102172099

    authors: Hagihara Y,Kim PS

    更新日期:2002-05-14 00:00:00

  • Deciphering the genetic architecture of variation in the immune response to Mycobacterium tuberculosis infection.

    abstract::Tuberculosis (TB) is a major public health problem. One-third of the world's population is estimated to be infected with Mycobacterium tuberculosis (MTB), the etiological agent causing TB, and active disease kills nearly 2 million individuals worldwide every year. Several lines of evidence indicate that interindividua...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1115761109

    authors: Barreiro LB,Tailleux L,Pai AA,Gicquel B,Marioni JC,Gilad Y

    更新日期:2012-01-24 00:00:00

  • Bmi1 is critical for lung tumorigenesis and bronchioalveolar stem cell expansion.

    abstract::Understanding the pathways that control epithelial carcinogenesis is vital to the development of effective treatments. The Polycomb group family member Bmi1 is overexpressed in numerous epithelial tumors, but its role in their development has not been established. We now show a key role for Bmi1 in lung adenocarcinoma...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0803574105

    authors: Dovey JS,Zacharek SJ,Kim CF,Lees JA

    更新日期:2008-08-19 00:00:00

  • Heat-shock protein hsp90 governs the activity of pp60v-src kinase.

    abstract::During or immediately after synthesis in vertebrate cells, the oncogenic protein-tyrosine kinase pp60v-src associates with the approximately 90-kDa heat-shock protein (hsp90). In this complex, pp60v-src is not functional as a kinase. When pp60v-src is subsequently found inserted into the plasma membrane, it is active ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.90.15.7074

    authors: Xu Y,Lindquist S

    更新日期:1993-08-01 00:00:00

  • The Epstein-Barr virus oncogene product, latent membrane protein 1, induces the downregulation of E-cadherin gene expression via activation of DNA methyltransferases.

    abstract::The latent membrane protein (LMP1) of Epstein-Barr virus (EBV) is expressed in EBV-associated nasopharyngeal carcinoma, which is notoriously metastatic. Although it is established that LMP1 represses E-cadherin expression and enhances the invasive ability of carcinoma cells, the mechanism underlying this repression re...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.152059399

    authors: Tsai CN,Tsai CL,Tse KP,Chang HY,Chang YS

    更新日期:2002-07-23 00:00:00

  • In vitro selection of optimal DNA substrates for T4 RNA ligase.

    abstract::We have used in vitro selection techniques to characterize DNA sequences that are ligated efficiently by T4 RNA ligase. We find that the ensemble of selected sequences ligated about 10 times as efficiently as the random mixture of sequences used as the input for selection. Surprisingly, the majority of the selected se...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.90.4.1576

    authors: Harada K,Orgel LE

    更新日期:1993-02-15 00:00:00

  • Calcium-dependent switching of the specificity of phosphoinositide binding to synaptotagmin.

    abstract::The synaptic vesicle membrane protein synaptotagmin (tagmin) is essential for fast, calcium-dependent, neurotransmitter release and is likely to be the calcium sensor for exocytosis, because of its many calcium-dependent properties. Polyphosphoinositides are needed for exocytosis, but it has not been known why. We now...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.93.23.13327

    authors: Schiavo G,Gu QM,Prestwich GD,Söllner TH,Rothman JE

    更新日期:1996-11-12 00:00:00

  • Low-barrier hydrogen bond in photoactive yellow protein.

    abstract::Low-barrier hydrogen bonds (LBHBs) have been proposed to play roles in protein functions, including enzymatic catalysis and proton transfer. Transient formation of LBHBs is expected to stabilize specific reaction intermediates. However, based on experimental results and theoretical considerations, arguments against th...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0811882106

    authors: Yamaguchi S,Kamikubo H,Kurihara K,Kuroki R,Niimura N,Shimizu N,Yamazaki Y,Kataoka M

    更新日期:2009-01-13 00:00:00

  • Observations and interpretation of x-ray absorption edges in iron compounds and proteins.

    abstract::X-ray absorption spectra near the Kalpha edge have been measured in various iron group compounds using the intense synchrotron radiation at the Stanford Synchrotron Research Project. In the cubic compounds KMF3 where M = Mn+2, Fe+2, Co+2, Ni+2, and Zn+2, well resolved lines were observed and assigned to the 1s leads t...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.73.5.1384

    authors: Shulman GR,Yafet Y,Eisenberger P,Blumberg WE

    更新日期:1976-05-01 00:00:00

  • RSK1 drives p27Kip1 phosphorylation at T198 to promote RhoA inhibition and increase cell motility.

    abstract::p90 ribosomal S6 kinase (RSK1) is an effector of both Ras/MEK/MAPK and PI3K/PDK1 pathways. We present evidence that RSK1 drives p27 phosphorylation at T198 to increase RhoA-p27 binding and cell motility. RSK1 activation and p27pT198 both increase in early G(1). As for many kinase-substrate pairs, cellular RSK1 copreci...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0805057106

    authors: Larrea MD,Hong F,Wander SA,da Silva TG,Helfman D,Lannigan D,Smith JA,Slingerland JM

    更新日期:2009-06-09 00:00:00

  • Controlling interactions in supported bilayers from weak electrostatic repulsion to high osmotic pressure.

    abstract::Understanding interactions between membranes requires measurements on well-controlled systems close to natural conditions, in which fluctuations play an important role. We have determined, by grazing incidence X-ray scattering, the interaction potential between two lipid bilayers, one adsorbed on a solid surface and t...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1211669109

    authors: Hemmerle A,Malaquin L,Charitat T,Lecuyer S,Fragneto G,Daillant J

    更新日期:2012-12-04 00:00:00

  • Isolation and characterization of the gene coding for cytosolic phosphoenolpyruvate carboxykinase (GTP) from the rat.

    abstract::The gene for cytosolic phosphoenolpyruvate carboxykinase (GTP) [GTP:oxaloacetate carboxy-lyase (transphosphorylating), EC 4.1.1.32] from the rat was isolated from a recombinant library containing the rat genome in phage lambda Charon 4A. The isolated clone, lambda PCK1, contains the complete gene for phosphoenolpyruva...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.80.12.3656

    authors: Yoo-Warren H,Monahan JE,Short J,Short H,Bruzel A,Wynshaw-Boris A,Meisner HM,Samols D,Hanson RW

    更新日期:1983-06-01 00:00:00

  • Adaptive evolution in the SRZ chemoreceptor families of Caenorhabditis elegans and Caenorhabditis briggsae.

    abstract::We investigated the possibility of positive selection acting on members of the putative seven-pass chemoreceptor superfamily in Caenorhabditis elegans, which comprises approximately 1,300 genes encoding seven-pass G protein-coupled receptors (GPCRs). Using a maximum-likelihood approach, we conducted statistical tests ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0406469102

    authors: Thomas JH,Kelley JL,Robertson HM,Ly K,Swanson WJ

    更新日期:2005-03-22 00:00:00

  • Adult hemoglobins are synthesized in erythroid colonies in vitro derived from murine circulating hemopoietic progenitor cells during embryonic development.

    abstract::Circulating peripheral blood cells and disaggregated yolk sac cells were obtained from normal mouse embryos as early as day 9 of gestation, prior to the formation of the fetal liver. These were cultured in vitro in plasma clots or methylcellulose, in the presence of either embryonic fluid or adult spleen cell-conditio...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.79.9.2952

    authors: Wong PM,Clarke BJ,Carr DH,Chui DH

    更新日期:1982-05-01 00:00:00

  • Global temperature responses to current emissions from the transport sectors.

    abstract::Transport affects climate directly and indirectly through mechanisms that cause both warming and cooling of climate, and the effects operate on very different timescales. We calculate climate responses in terms of global mean temperature and find large differences between the transport sectors with respect to the size...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0804844105

    authors: Berntsen T,Fuglestvedt J

    更新日期:2008-12-09 00:00:00

  • Biophysical and biological activity of a synthetic 8.7-kDa hydrophobic pulmonary surfactant protein SP-B.

    abstract::We have synthesized pulmonary surfactant apoprotein SP-B peptides by solid-phase chemistry and demonstrated their ability to enhance the surface-active properties of synthetic lipid mixtures. The synthetic peptides were reactive with antiserum generated against the native bovine surfactant peptide. Both peptides confe...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.87.7.2633

    authors: Sarin VK,Gupta S,Leung TK,Taylor VE,Ohning BL,Whitsett JA,Fox JL

    更新日期:1990-04-01 00:00:00