2 edition of Targeted gene knockout of Tetrahymena dynein heavy chain gene DYH13 found in the catalog.
Targeted gene knockout of Tetrahymena dynein heavy chain gene DYH13
Jonelle Ruth Zimmerman
Written in English
|Statement||by Jonelle Ruth Zimmerman|
|The Physical Object|
|Pagination||iii, 55 leaves :|
|Number of Pages||55|
approach to targeted gene knockout in mammalian cells, using engineered zinc-ﬁnger nucleases (ZFNs). ZFNs can be designed to target a chosen locus with high speciﬁcity. Upon transient expres-sion of these nucleases the target gene is ﬁrst cleaved by the ZFNs and then repaired by a natural—but imperfect—DNA repair pro-File Size: KB. This gene encodes the catalytic subunit of the DNA-dependent protein kinase (DNA-PK). It functions with the Ku70/Ku80 heterodimer protein in DNA double strand break repair and recombination. The protein encoded is a member of the PI3/PI4-kinase family.[provided by RefSeq, Jul ].
2 Overview: Tetrahymena as a valuable genetic unicellular animal model Tetrahymena thermophila belongs to the Alveolates, a major evolutionary branch of eukaryotic protists composed of three primary lineages: Ciliates (e.g., Tetrahymena and Paramecium), Dinoflagellates (e.g., Symbiodinium, the coral endosymbiont, and Alexandrium, which causes paralytic shellfish poisoning). 1 Tetrahymena Comparative Genomics Sequencing Project A Revised White Paper Submitted to the NHGRI Comparative Genome Evolution Working Group Ma Summary of Request Tetrahymena thermophila is by far the most well-developed model organism for basic and biomedical research among the entire Alveolate clade, a major and diverse.
Targeted gene knockout by direct delivery of ZFN proteins Thomas Gaj, 1 Jing Guo, 1 Yoshio Kato, 1, 2 Shannon J. Sirk, 1 and Carlos F. Barbas, III 1 1 The Skaggs Institute for Chemical Biology and the Departments of Molecular Biology and Chemistry, The Scripps Research Institute, La Jolla, California, USA. to each heavy chain obtained by the densitometry of a Coomassie-stained gel was , , for fractions a, c, and d, respectively. These values are similar to those obtained for Chlamydomonas inner-arm dyneins (our un- published data). 4. Diiussion We have shown that the 14s dynein of Tetrahymena.
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Targeted gene knockout of inner arm 1 in Tetrahymena thermophila Steven P. Angus a, Richard E. Edelmann b, David G. Pennock a1) a Department of Zoology, Miami University, Oxford, OH/USA b Department of Botany, Miami University, Oxford, OH/USA Received J Received in revised version March 9, Accepted March 9, Cilia – Tetrahymena – dynein – axoneme – cell Cited by: Targeted gene disruption of dynein heavy chain 7 of Tetrahymena thermophila results in altered ciliary waveform and reduced swim speed.
dynein hea vy chain 7 protein (Dyh7p), a knockout. The targeted disruption of somatic genes in Tetrahymena thermophilapresents the opportunity to determine the contributions of individual dynein isoforms in a single cell that expresses multiple dynein heavy chain genes.
Substantial portions of twoTetrahymena cytoplasmic dynein heavy chain genes were cloned, and their motor domains were sequenced. The criteria for predicting introns are based on our experience with the 64 introns that we experimentally confirmed in our studies of the Tetrahymena cytoplasmic dynein heavy chains 1 and 2 and the axonemal dynein β heavy chain (Lee et al.,Lincoln et al., ).
The introns are 50– bp in length (average = 80 bp) and have –95 Cited by: 3. THE ciliated protozoan Tetrahymena thermophila can grow at an exceptionally high rate (its doubling time is approximately 2 hr) and can reach a high density (a few million cells per milliliter) under simple and inexpensive culture conditions (reviewed in Orias et al.
).In combination with robust genetic manipulation methods (reviewed in Chalker ), Tetrahymena is a useful model Cited by: At least 21 mutations in the DNAI1 gene have been found to cause primary ciliary dyskinesia, which is a condition characterized by respiratory tract infections, abnormal organ placement, and an inability to have children (infertility).
DNAI1 gene mutations result in an absent or abnormal intermediate chain 1. Without a normal version of this subunit, the ODAs cannot form properly and.
Abstract. In this chapter, we outline methods to modify targeted genes in a strategy could be utilized to deduce the function of individual dynein isoforms, as well as to dissect the functional domains within a single heavy by: 2.
Targeted gene knockout by direct delivery of ZFN proteins Thomas Gaj, 1 Jing Guo, 1 Yoshio Kato, 1, 2 Shannon J. Sirk, 1 and Carlos F. Barbas, III 1 1 The Skaggs Institute for Chemical Biology and the Departments of Molecular Biology and Chemistry, The Scripps Research Institute, La Cited by: Tetrahymena is a genus of free-living ciliates that can also switch from commensalistic to pathogenic modes of survival.
They are common in freshwater ponds. They are common in freshwater ponds. Tetrahymena species used as model organisms in biomedical research are Class: Oligohymenophorea. The construct often is engineered with the target-gene sequence disrupted by the neoR gene and with the tkHSV gene at one end, beyond the sequence of the target gene.
Most constructs will insert at random by nonhomolo-gous recombination rather than by gene-targeted insertion through homologous recombination. (gene) in the plasmid greatly increases the frequency of recombination both in yeast (Orr-Weaver et al., ) and mammalian (Smithies et cells al., ).
Moreover, the DSB targeted the exogenous. Gene knockout (GO) is a genetic technique supplemented with biotechnological tool, in which an organism is engineered to carry gene that has been made inoperative. These genes are known as knockouts; used in assigning function to specific genes having unknown function.
Gene knockout strategy, reverse genetic tools, used to determine the function of target genes by gene technology. Existing methods for gene disruption are limited by their efficiency, time to completion, and/or the potential for confounding off-target effects.
Here, we demonstrate a rapid single-step approach to targeted gene knockout in mammalian cells, using engineered zinc-finger nucleases (ZFNs). Current gene therapy protocols against cancer often have limited target specificity.
Here, a novel tumor-specific targeted gene delivery procedure, which is based on Tetrahymena group I intron ribozyme, is presented.
This ribozyme can target a cancer-specific transcript and then replace the RNA with new transcripts, resulting in induction of the transgene activity selectively in cancer cells Cited by: What's New. Tetrahymena thermophila macronuclear genome fully completed.
Posted on by ciliate-curator. We have received an update of the Tetrahymena thermophila genome sequence from researchers at Ocean University of China.
The gene pages, BLAST server, and Genome Browser have all been updated accordingly to display these new data. targeted gene knockout by direct delivery of zinc-finger nuclease proteins. Thomas Gaj. 1–3, Jing Guo. 1–3, Yoshio Kato. 1–4, Shannon J Sirk. 1–3 & Carlos F Barbas III. 1–3. Zinc-finger nucleases (Zfns) are versatile reagents that have redefined genome engineering.
realizing the full potential of. Well the simplest way is to cut the gene at the both ends from operon by means of proper restriction enzymes but in case that you get more than one recognition site then you can use microRNA or proper oligo to make a hairpin loop and break that part If you only want to silence the operon then you just need to cut it nearly at middle of the gene and ligate it with a nonsense sequence, then it.
When cells that have loxP sites in their genome express Cre, a recombination event can occur between the loxP sites. Cre recombinase binds to the first and last 13 bp regions of a lox site forming a dimer. This dimer then binds to a dimer on another lox site to form a sites are directional and the two sites joined by the tetramer are parallel in orientation.
Identifies gene interaction in which a gene at one locus mask (suppress)the effects of a gene at a different locus What is genetic complementation Assay Generate multiple mutants w/ same pheno, to identify multiple genes in a particular pathway.
Gene-symbol tm#c(project abbreviation)Labcode is created when the original ‘tm1a’ allele undergoes flp-mediated recombination, which removes the LacZ reporter, a loxP site, and neo, leaving an intact gene with loxP flanking the critical exon.
This allele can then be used for future cre-mediated conditional mutagenesis. The allele symbol has a superscripted portion comprised of. Nitin Puri, Alokes Majumdar, Bernard Cuenoud, Francois Natt, Pierre Martin, Andre Boyd, Paul S.
Miller, Michael M. Seidman.The Tetrahymena genome size (roughly Megabase pairs) is of the same order of magnitude as that of Drosophila-one order larger than yeast (Saccharomyces) and one smaller than human.
Figure 1. Scanning electron micrograph of a Tetrahymena cell. ***** Tetrahymena's versatile biology provides extremely useful experimental tools.Indeed, using the ZFN methodology, targeted gene integration frequencies of 15, 6 and 5% have been observed, respectively, for insertion of a 12 bp tag, a bp open reading frame and a kb promoter-transcription unit at a specific location in human cells, without selection for desired ed Gene Integration.
7 In addition.