Abstract:
:Emerging evidence suggests that recombinant adeno-associated viral (rAAV) vectors can be used for specific gene targeting in human somatic cells. We have developed an rAAV vector construction procedure employing fusion PCR and a single cloning step that considerably simplifies the knockout process. We demonstrate its utility by disrupting genes at specific positions within human colon cancer cells as well as within immortalized normal epithelial cells. This technology should be broadly applicable to in vitro studies that require the manipulation of the human genome.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Kohli M,Rago C,Lengauer C,Kinzler KW,Vogelstein Bdoi
10.1093/nar/gnh009keywords:
subject
Has Abstractpub_date
2004-01-02 00:00:00pages
e3issue
1eissn
0305-1048issn
1362-4962pii
32/1/e3journal_volume
32pub_type
杂志文章abstract::Interest in secondary metabolites such as RiPPs (ribosomally synthesized and posttranslationally modified peptides) is increasing worldwide. To facilitate the research in this field we have updated our mining web server. BAGEL4 is faster than its predecessor and is now fully independent from ORF-calling. Gene clusters...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky383
更新日期:2018-07-02 00:00:00
abstract::siRNAs confer sequence specific and robust silencing of mRNA. By virtue of these properties, siRNAs have become therapeutic candidates for disease intervention. However, their use as therapeutic agents can be hampered by unintended off-target effects by either or both strands of the siRNA duplex. We report here that u...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq961
更新日期:2011-03-01 00:00:00
abstract::Human 2'-5' oligoadenylate synthetase-1 (OAS1) is central in innate immune system detection of cytoplasmic double-stranded RNA (dsRNA) and promotion of host antiviral responses. However, the molecular signatures that promote OAS1 activation are currently poorly defined. We show that the 3'-end polyuridine sequence of ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gku1289
更新日期:2015-01-01 00:00:00
abstract::Pyrimidine adducts in cellular DNA arise from modification of the pyrimidine 5,6-double bond by oxidation, reduction or hydration. The biological outcome includes increased mutation rate and potential lethality. A major DNA N:-glycosylase responsible for the excision of modified pyrimidine bases is the base excision r...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/29.3.604
更新日期:2001-02-01 00:00:00
abstract::All exchangeable protons in a short DNA helix, d(CG)3 sodium salt, have been studied by proton nuclear magnetic resonance. The cytidine and guanosine amino protons have been assigned for the first time. As a function of temperature the cytidine amino protons and the imino protons behave very similarly, their relaxatio...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.21.8269
更新日期:1984-11-12 00:00:00
abstract::The adenovirus late region 1 (L1) represents an example of an alternatively spliced gene where one 5' splice site is spliced to two alternative 3' splice sites, to produce two mRNAs; the 52,55K and IIIa mRNAs, respectively. Accumulation of the L1 mRNAs is temporally regulated during the infectious cycle. Thus, the pro...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/22.3.332
更新日期:1994-02-11 00:00:00
abstract::In non-malignant RWPE-1 prostate epithelial cells signaling by the nuclear receptor Vitamin D Receptor (VDR, NR1I1) induces cell cycle arrest through targets including CDKN1A (encodes p21((waf1/cip1))). VDR dynamically induced individual histone modification patterns at three VDR binding sites (R1, 2, 3) on the CDKN1A...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq875
更新日期:2011-03-01 00:00:00
abstract::Thousands of eukaryotic protein-coding genes generate circular RNAs that have covalently linked ends and are resistant to degradation by exonucleases. To prove their circularity as well as biochemically enrich these transcripts, it has become standard in the field to use the 3'-5' exonuclease RNase R. Here, we demonst...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz576
更新日期:2019-09-19 00:00:00
abstract::During DNA replication replicative polymerases move in discrete mechanical steps along the DNA template. To address how the chemical cycle is coupled to mechanical motion of the enzyme, here we use optical tweezers to study the translocation mechanism of individual bacteriophage Phi29 DNA polymerases during processive...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv204
更新日期:2015-04-20 00:00:00
abstract::The discoidin proteins of Dictyostelium discoideum are highly expressed during development. The Disc I gamma promoter allows the regulation of heterologous protein expression by experimental conditions. We report conditions under which the promoter activity is efficiently repressed during growth in the wildtype strain...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/21.6.1397
更新日期:1993-03-25 00:00:00
abstract::Ribosomal RNA (rRNA) in intact ribosomes is cleaved into discrete products on incubation of reticulocyte lysates or L-cell extracts with ppp(A2'p)3A. Cleavage of rRNA may, therefore, provide a useful assay for 2-5A (ppp)A2'p)nA; n = 2 to 4) or for the presence of a 2-5A-dependent nuclease. The results with reticulocyt...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/9.7.1571
更新日期:1981-04-10 00:00:00
abstract::Type II restriction endonucleases Bam HI and Eco RI were covalently coupled to Sepharose. These insolubilized enzymes generated fragment patterns for several viral DNAs identical to those produced by the respective free enzymes. Conditions for optimal activity were similar for both bound and unbound forms of the enzym...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/5.3.679
更新日期:1978-03-01 00:00:00
abstract::The 'regions' method for multisequence alignment used in the previously reported program MALIGN has been generalized to include recursive refinement so that unaligned portions between two regions at the current level of resolution can be handled with increased resolution. Additionally, there is incorporated a limiting...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/16.5.1683
更新日期:1988-03-11 00:00:00
abstract::We have isolated the poly(A) polymerase (PAP) encoding gene pla1 [for poly(A) polymerase] from the fission yeast Schizosaccharomyces pombe. Protein sequence alignments with other poly(A) polymerases reveal that pla1 is more closely related to Saccharomyces cerevisiae PAP than to bovine PAP. The two yeast poly(A) polym...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/24.13.2585
更新日期:1996-07-01 00:00:00
abstract::The thermal stabilities of RNA:DNA hybrids are substantially greater than those of DNA:DNA duplexes in aqueous electrolyte solutions containing high concentrations of formamide. Association rates to form DNA:DNA duplexes and DNA:RNA hybrids have been measured in these solvents. There is a temperature range in which DN...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/4.5.1539
更新日期:1977-01-01 00:00:00
abstract::In Saccharomyces cerevisiae the majority of the genes for 5S rRNA lie within a 9kb rDNA sequence that is present as 100-200 tandemly-repeated copies on Chromosome XII. Following our observations that about 10% of yeast 5S rRNA exists as minor variant sequences, we screened a collection of yeast DNA fragments cloned in...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.10.4083
更新日期:1984-05-25 00:00:00
abstract::The satellite RNA of tobacco ringspot virus (STobRV RNA) replicates and becomes encapsidated in association with tobacco ringspot virus. Previous results show that the infected tissue produces multimeric STobRV RNAs of both polarities. RNA that is complementary to encapsidated STobRV RNA, designated as having the (-) ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/14.24.9729
更新日期:1986-12-22 00:00:00
abstract::The nature of mammalian origins of DNA replication remains controversial and this is primarily because two-dimensional gel replicon mapping techniques have identified broad zones of replication initiation whereas several other techniques, such as quantitative PCR, have disclosed more discrete sites of initiation at th...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.10.2313
更新日期:1998-05-15 00:00:00
abstract::The European Bioinformatics Institute (EMBL-EBI) supports life-science research throughout the world by providing open data, open-source software and analytical tools, and technical infrastructure (https://www.ebi.ac.uk). We accommodate an increasingly diverse range of data types and integrate them, so that biologists...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx1154
更新日期:2018-01-04 00:00:00
abstract::Telomeres play a central role in human cancer, cardiovascular aging and possibly longevity. However, present methods to measure telomere length are fraught with shortcomings that limit their use. Here, we describe a novel method to measure the relative telomere DNA content by dot blot analysis. In each dot, the DNA co...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr235
更新日期:2011-07-01 00:00:00
abstract::A cDNA (2855 nt) encoding a putative cytotoxic ribonuclease (rapLR1) related to the antitumor protein onconase was cloned from a library derived from the liver of gravid female amphibian Rana pipiens. The cDNA was mainly comprised (83%) of 3' untranslated region (UTR). Secondary structure analysis predicted two unusua...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/28.12.2375
更新日期:2000-06-15 00:00:00
abstract::Chlorella virus DNA ligase is the smallest eukaryotic ATP-dependent DNA ligase known; it suffices for yeast cell growth in lieu of the essential yeast DNA ligase Cdc9. The Chlorella virus ligase-adenylate intermediate has an intrinsic nick sensing function and its DNA footprint extends 8-9 nt on the 3'-hydroxyl (3'-OH...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkg665
更新日期:2003-09-01 00:00:00
abstract::In testis mRNA stability and translation initiation are extensively under the control of poly(A)-binding proteins (PABP). Here we have cloned a new human testis-specific PABP (PABP3) of 631 amino acids (70.1 kDa) with 92.5% identical residues to the ubiquitous PABP1. A northern blot of multiple human tissues hybridise...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/29.9.1872
更新日期:2001-05-01 00:00:00
abstract::The third intron from Physarum polycephalum (Pp LSU 3) is one of the closest known relatives to the well-studied Tetrahymena group I intron. Both introns are located at the same position in the 26S rRNA gene, and with the exception of an open reading frame in Pp LSU 3, are highly homologous. While Pp LSU 3 has been sh...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/22.20.4315
更新日期:1994-10-11 00:00:00
abstract::Sequence specific DNA methylation sometimes results in the protection of some or all of a restriction endonucleases' cleavage sites. This is usually, but not always, the result of methylation of one or both strands of DNA at the site characteristic of the corresponding "cognate" modification methylase. The known effec...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/9.22.5859
更新日期:1981-11-25 00:00:00
abstract::Exposure to many endogenous and exogenous agents can give rise to DNA alkylation, which constitutes a major type of DNA damage. Among the DNA alkylation products, alkyl phosphotriesters have relatively high frequencies of occurrence and are resistant to repair in mammalian tissues. However, little is known about how t...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky140
更新日期:2018-05-04 00:00:00
abstract::Alternative splicing is the primary source of proteome complexity in metazoans and its regulation shapes the proteome in response to shifting physiological requirements. We developed a bichromatic splicing reporter that uses a peculiar feature of some fluorescent protein coding regions to express two different fluores...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl967
更新日期:2006-01-01 00:00:00
abstract::Four pseudorevertants of a -10 region lacP mutation were isolated. Three of these mutations were found to activate nascent promoters. These mutations were: a -2 G/C----A/T change (-2A) promoting transcription at position +11, a +1 A/T----T/A change (+1T) promoting transcription initiation at position +13, and a +10 C/...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/17.10.3927
更新日期:1989-05-25 00:00:00
abstract::Measuring minimal residual disease in cancer has applications for prognosis, monitoring treatment and detection of recurrence. Simple sequence-based methods to detect nucleotide substitution variants have error rates (about 10-3) that limit sensitive detection. We developed and characterized the performance of MASQ (m...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa090
更新日期:2020-04-17 00:00:00
abstract::The Biodegradative Strain Database (BSD) is a freely-accessible, web-based database providing detailed information on degradative bacteria and the hazardous substances that they degrade, including corresponding literature citations, relevant patents and links to additional web-based biological and chemical data. The B...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkg032
更新日期:2003-01-01 00:00:00