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    Sunday, 03 February 2013 00:00

    Mutant Gene Gives Pigeons Fancy Hairdos

    DECODED GENOME REVEALS SECRETS OF PIGEON TRAITS AND ORIGINS

    Old Dutch capuchine manecrest2University of Utah researchers decoded the genetic blueprint of the rock pigeon, unlocking secrets about pigeons’ Middle East origins, feral pigeons’ kinship with escaped racing birds, and how mutations give pigeons traits like a fancy feather hairdo known as a head crest.

    “Birds are a huge part of life on Earth, and we know surprisingly little about their genetics,” especially compared with mammals and fish, says Michael D. Shapiro, one of the study’s two principal authors and an assistant professor of biology at the University of Utah. “There are more than 10,000 species of birds, yet we know very little about what makes them so diverse genetically and developmentally.”

    He adds that in the new study, “we’ve shown a way forward to find the genetic basis of traits – the molecular mechanisms controlling animal diversity in pigeons. Using this approach, we expect to be able to do this for other traits in pigeons, and it can be applied to other birds and many other animals as well.”

    The study appears Jan. 31 on Science Express, the website of the journal Science. Shapiro led the research with Jun Wang of China’s BGI-Shenzhen (formerly Beijing Genomics Institute) and other scientists from BGI, the University of Utah, Denmark’s University of Copenhagen and the University of Texas M.D. Anderson Cancer Center in Houston.

    Key findings of the study of pigeons, which first were domesticated some 5,000 years ago in the Mediterranean region:

    – The researchers sequenced the genome, or genetic blueprint, of the rock pigeon, Columba livia, among the most common and varied bird species on Earth. There are some 350 breeds with different sizes, shapes, colors, color patterns, beaks, bone structure, vocalizations and arrangements of feathers on the feet and head – including head crests that come in shapes known as hoods, manes, shells and peaks.

    The pigeon is among the few bird genomes sequenced so far, along with those of the chicken, turkey, zebra finch and a common parakeet known as a budgerigar or budgie, so “this will give us new insights into bird evolution,” Shapiro says.

    – Using innovative software developed by study co-author Mark Yandell, a University of Utah professor of human genetics, the scientists revealed that a single mutation in a gene named EphB2 causes head and neck feathers to grow upward instead of downward, creating head crests.

    “This same gene in humans has been implicated as a contributor to Alzheimer’s disease as well as prostate cancer and possibly other cancers,” Shapiro says, noting that more than 80 of the 350 pigeon breeds have head crests, which play a role in attracting mates in many bird species.

    – The researchers compared the pigeon genome to those of chickens, turkeys and zebra finches. “Despite 100 million years of evolution since these bird species diverged, their genomes are very similar,” Shapiro says.

    – The study turned up more conclusive evidence that major pigeon breed groups originated in the Middle East, and that North American feral pigeons – which are free-living but not wild – are close relatives of racing pigeons, named racing homers.

    A Genome for the Birds, a Gene for Head Crests

    The study assembled 1.1 billion base pairs of DNA in the rock pigeon genome, and the researchers believe there are about 1.3 billion total, compared with 3 billion base pairs in the human genome. The rock pigeon’s 17,300 genes compare with about 21,000 genes in people.

    The researchers first constructed a “reference genome” – a full genetic blueprint – from a male of the pigeon breed named the Danish tumbler. They did less complete sequencing of two feral pigeons and 38 other pigeons from 36 breeds.

    Shapiro says his team’s study is the first to pinpoint a gene mutation responsible for a pigeon trait, in this case, head crests.

    “A head crest is a series of feathers on the back of the head and neck that point up instead of down,” Shapiro says. “Some are small and pointed. Others look like a shell behind the head; some people think they look like mullets. They can be as extreme as an Elizabethan collar.”

    The study found strong evidence that the EphB2 (Ephrin receptor B2) gene acts like an on-off switch to create a head crest when mutant, and no head crest when normal. It also showed the mutation and related changes in nearby DNA are shared by all crested pigeons, so the trait evolved just once and was spread to numerous pigeon breeds by breeders. They ruled out the alternate possibility the mutation arose several times independently in different breeds.

    The researchers analyzed full or partial genetic sequences for 69 crested birds from 22 breeds, and 95 uncrested birds from 57 breeds. They found a perfect association between the mutant gene and the presence of head crests.

    “The way we tracked this trait was innovative,” Shapiro says. “We used gene-finding software from Mark Yandell’s group that was developed to find mutations that control human diseases. We adapted this software to find mutations that control interesting traits in pigeons. This should be extendable to other animals as well.”

    The scientists also showed that while the head crest trait becomes apparent in juvenile pigeons, the mutant gene affects pigeon embryos by reversing the direction of feather buds – from which feathers later grow – at a molecular level.

    Other genetic factors – not identified in the new study – determine what kind of head crest east pigeon develops: shell, peak, mane or hood, according to Shapiro.

    Tracking the Origins of Pigeons

    A 2012 by Shapiro study provided limited evidence of pigeons’ origins in the Middle East and some breeds’ origins in India, and indicated kinship between common feral or free-living city pigeons and escaped racing pigeons.

    In the new study, “we included some different breeds that we didn’t include in the last analysis,” Shapiro says. “Some of those breeds only left the Middle East in the last few decades. They’ve probably been there for hundreds if not thousands of years. If we find that other breeds are closely related to them, then we can infer those other breeds probably also came from the Middle East. That’s what we did.”

    “We found that the owl breeds – which are pigeon breeds with very short beaks and that are very popular with breeders – likely came from the Middle East,” he says. “They are very closely related to breeds we know came from Syria, Lebanon and Egypt.”

    Shapiro says the study also “found a lot of shared genetic heritage between breeds from Iran and breeds we suspect are from India, consistent with historical records of trade routes between those regions. People were not only trading goods along those routes, but probably also interbreeding their pigeons.”

    As for the idea that free-living pigeons descended from escaped racing pigeons, Shapiro says his 2012 study was based on “relatively few genetic markers scattered throughout the genome. We now have stronger evidence based on 1.5 million markers, confirming the previous result with much better data.”

    The scientists analyzed partial genomes of two feral pigeons: one from a U.S. Interstate-15 overpass in the Salt Lake Valley, and the other from Lake Anna in Virginia.

    “Despite being separated by 1,000 miles, they are genetically very similar to each other and to the racing homer breed,” Shapiro says.

    He notes that pigeons were one of evolutionist Charles Darwin’s “favorite examples of how selection works. He used this striking example of artificial selection [by breeding] to communicate how natural selection works. Now we can get to the DNA-level changes that are responsible for some of the diversity that intrigued Darwin 150 years ago.”

    The study’s University of Utah co-authors were Yandell; Eric Domyan, biology postdoctoral fellow; Zev Kronenberg and Michael Campbell, Ph.D. students in human genetics; Anna Vickery, biology undergraduate student; Sydney Stringham, Ph.D. student in biology; and Chad Huff, a former postdoctoral fellow in human genetics now at the University of Texas.

    The study was funded by the Burroughs Wellcome Fund, the National Science Foundation, the University of Utah Research Foundation, the National Institutes of Health and the Danish National Research Foundation.

    Published in News
    Wednesday, 06 March 2013 15:50

    Targatt transgenic Kit

    TARGATT Transgenic Kit is designed to create site-specific, knock-in transgenic mice in a more efficient and significantly faster way over traditional methods. Generating transgenic mice by conventional methods (e.g. pronuclear microinjection or lentiviral injection) has several limitations, first of which is random insertion of the transgene. Random insertion of a transgene results in a position effect where either the transgene is prone to silencing or endogenous gene expression is disrupted. Furthermore, this position effect is compounded when a transgene is inserted as multiple copies, resulting in instability at the insertion locus.

    Using our proprietary site-specific DNA integration system, TARGATT Transgenic Kit, combined with our genetically TARGATT mouse embryos (cat.# AST-0001, AST-0002, AST-0003, AST-0004). Targatt offers high quality H11(Hipp11) and ROSA26 transgenic mouse kits with our TARGATT technology. Using our TARGATT Technology, you can generate your own knock-in mouse in just three months.

    targatt-antibodies-gentaur-knockin-knockout-mouse-1

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    If you prefer to generate a knock-in mouse model on your own, TARGATT Embryo H11(Hipp11) or Rosa26 and Transgenic Kits (2 or 5 microinjection size) are available for purchase.

    Background:

    TARGATT technology enables highly efficient site-specific gene integration in mammalian cells and animals1, 2. This technology uses øC31 integrase to insert any gene of interest into a docking site, pre-engineered in an intergenic region and transcriptionally active genomic locus. Our TARGATT technology improves several aspects in the generation of transgenic cell lines and animals: (1) High integration efficiency mediated by øC31 integrase reduces time and cost; (2) Site-specific integration at a pre-selected genomic locus eliminates position effect and ensures high level expression of the transgene; (3) Integration at intergenic region ensures that no internal genes are interrupted; (4) Single copy gene integration eliminates repeat-induced gene silencing and genomic instability; (5) Site-specific integration allows a precise comparison of the effects of the transgenes among different lines. TARGATT technology can be utilized for a variety of applications including reporter gene expression, gene knockdown, disease cell and animal models.

    targatt-antibodies-gentaur-knockin-knockout-mouse-3

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    Published in Promos
    Wednesday, 06 March 2013 15:35

    TARGATT embryos

    TARGATT embryos are derived from one of our genetically engineered mouse models. These mouse embryos can be used as embryo donors for creating site-specific transgenic mice in a more efficient and faster way compared to traditional methods. Generating transgenic mice by conventional methods (e.g. pronuclear microinjection or lentiviral injection) has several limitations, first of which is the random insertion of the transgene. Random insertion of a transgene results in a position effect where either the transgene is prone to silencing or endogenous gene expression is disrupted. Furthermore, this position effect is compounded when transgenes are inserted in multiple copies, resulting in genomic instability at the insertion locus. Using our proprietary site-specific DNA integration technology, TARGATT embryos combined with our TARGATT Transgenic Kit (cat# AST-1001 or AST-1002), you can generate your desired transgenic mouse models with guaranteed gene expression faster.

    Gentaur offers high quality H11 and ROSA26 TARGATT frozen embryos. Using our TARGATT Technology, you can generate your own knock-in mouse from H11 or ROSA26 embryos in 3 months. Our H11(Hipp11) and ROSA26 frozen embryos contain 3 straws with a total 45-60 cryopreserved embryos (8-cell stage/morula). They are shipped in a dry shipper containing liquid nitrogen. We also offer fresh H11 and ROSA26 embryos with each vial containing 25-35 fresh embryos (E3.5) in KSOM medium. We offer overnight shipping at ambient temperature. Embryos will be transferred to recipient(s) immediately upon arrival. US customers only.

    targatt-antibodies-gentaur-knockin-knockout-mouse-1

    targatt-antibodies-gentaur-knockin-knockout-mouse-2

    If you prefer to generate a knock-in mouse model on your own, TARGATT Embryo (H11 or Rosa26) and Transgenic Kits (2 or 5 microinjection size) are available for purchase.

    Background:

    TARGATT technology enables highly efficient site-specific gene integration in mammalian cells and animals1, 2. This technology uses øC31 integrase to insert any gene of interest into a docking site, pre-engineered in an intergenic region and transcriptionally active genomic locus. Our TARGATT technology improves several aspects in the generation of transgenic cell lines and animals: (1) High integration efficiency mediated by  øC31 integrase reduces time and cost; (2) Site-specific integration at a pre-selected genomic locus eliminates position effect and ensures high level expression of the transgene; (3) Integration at intergenic region ensures that no internal genes are interrupted; (4) Single copy gene integration eliminates repeat-induced gene silencing and genomic instability; (5) Site-specific integration allows a precise comparison of the effects of the transgenes among different lines. TARGATT technology can be utilized for a variety of applications including reporter gene expression, gene knockdown, disease cell and animal models.

    targatt-antibodies-gentaur-knockin-knockout-mouse-3

    Click here to see all products

    Published in Promos
    Wednesday, 06 March 2013 11:03

    TARGATT Knockin Mice - Stem Cells Products

    targatt-stemcells-knockin-mouse-1TARGATT Embryos

    Using our novel TARGATT system, a gene of interest can be specifically inserted at a well-characterized, transcriptionally-active locus in the mouse genome with guaranteed transgene expression. Tissue-specific and / or ubiquitous expression options are available.

     

    Advantages of TARGATT technology:

    1. Site-specific gene integration at a transcriptionally-active locus ensures high-level gene expression.
    2. Integration happens at an intergenic region; no internal genes are disrupted.
    3. The integrase system catalyzes a unidirectional integration event and results in a high efficiency in producing transgenic mice.
    4. Gene integration at the same locus allows a precise comparison of the transgenics from one line to another.

     

    targatt-stemcells-knockin-mouse-2TARGATT Kits

    TARGATT Supporting Materials

     

     

    Mouse Embryonic Fibroblasts (MEF)

    targatt-stemcells-knockin-mouse-3  CF-1
      Neo-resistant
      DR4
      SNL (STO feeder cells)

    SNL (STO feeder cells)

     

    Cell Culture Products

      Germline-tested & ESC-qualified FBS
      Specialty Media
      Basal Media (DMEM)
      Stem Cell Growth and Differentiation Factors
      ASC Small Molecules
      3D Culture and Expansion System

    Reprogramming

    targatt-stemcells-knockin-mouse-4

    targatt-stemcells-knockin-mouse-5

     

     

     

     

     

     

     

    ESC/iPSC Characterization

      Pluripotency Protein Markers                   Stem Cell Gene Array
     targatt-stemcells-knockin-mouse-6    targatt-stemcells-knockin-mouse-7

    ●  Pluripotency mRNA Markers
    ●  Components

     

    ESC/iPSC Differentiation

      Neural Differentiation
      Dendritic Cell (DC) Generation

    ES/iPS Cell Lines

    Mouse ES Cell Lines                                 Human iPS Cells

    targatt-stemcells-knockin-mouse-8   targatt-stemcells-knockin-mouse-9

    Cell Depository

    Primary Cell, cDNA, RNA (Normal)

    targatt-stemcells-knockin-mouse-10  Endocrine
      Neural
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      Urinary
      Reproductive
      Skeletal Muscle
      Hematopoietic
      Integumentary
      Cardiovascular
      Miscellaneous

    Primary Cell, cDNA, RNA (Disease)

      Blood Disorders
      Neurological Disorders
      Degenerative Disorders
      Metabolic Disorders
      Cardiovascular Disorders
      Congenital Disorders
      Endocrine Disorders
      Autoimmune Disorders
      Genetic Disorders
      Muscular Disorders
      Oncogenic Disorders

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    Read more about Targatt gene modification here

    Published in Promos
    Monday, 28 January 2013 11:36

    TARGATT Gene Modification

    TARGATT Fast & Efficient Gene Modification in Human Cells. Make your cell line in 3 months!

    Based on our proprietary site-specific TARGATT technology for fast knock-in mouse (KI) generation, Applied StemCell Inc. (ASC) recently developed a TARGATT system specifically for introducing gene(s) of interest into mammalian cell lines. The unique advantage of this system is that any gene of interest can be inserted efficiently into a defined, transcriptionally-active locus with high gene expression in a single copy fashion.

    targat talen knockin celladvantage

    targatt talen knockout genemodification

    Technical Details:

    TARGATT technology enables highly efficient site-specific gene integration in mammalian cells and animals1, 2. This technology uses ?C31 integrase to insert any gene of interest into a docking site, pre-engineered in an intergenic region and transcriptionally active genomic locus. Our TARGATT technology improves several aspects in the generation of transgenic cell lines and animals: (1) High integration efficiency mediated by ?C31 integrase reduces time and cost; (2) Site-specific integration at a pre-selected genomic locus eliminates position effect and ensures high level expression of the transgene; (3) Integration at intergenic region ensures that no internal genes are interrupted; (4) Single copy gene integration eliminates repeat-induced gene silencing and genomic instability; (5) Site-specific integration allows a precise comparison of the effects of the transgenes among different lines. TARGATT technology can be utilized for a variety of applications including reporter gene expression, gene knockdown, disease cell and animal models.

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    knockin knockout targatt talen

    More information about Stem cells

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    Published in Promos