Can restriction enzymes be used to treat huntington disease?
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Sure it WOULD work if there were means to deliver it to each cell, ensure 100% restriction efficiency & specificity, and ligate the DNA nick afterwards. So far it is sci-fi; prenatal diagnostics is the way to go for now.
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Biotechnology covers a wide range of uses, but I'd say that mostly it's about manipulating cells to express genes that we want them to express.  This could be to make them produce industrial enzymes, designer agriculture, or biopharmaceuticals.  the genetic code is pretty much universal, meaning we can take a gene from one organiand put into another to acheive specific results.  Industrial enzymes can be used for anything from cleaning wastewater with special enzymes to degrade certain toxins, to making enzymes that breakdown starch into sugar for the production of ethanol for fuel, to the making of specific detergents.  With agriculture, we can transfect plants with genes that make them more resistant to diseases or vermin, or more robust and able to grow under more varied claimates, or we can make food bigger or more nutritious.  In biopharmceuticals, we address genetic disorders by creating the proteins that some people are unable to make for themselves, allowing them to lead more normal lives.
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You have to identify two sequences that are >6kb apart from one another that flank the sequence you are attempting to amplify. Better yet, you can get you a well understood cloning vector (plasmid) that already has primers develped for it that flank the polyrestriction site region (ie. puc19 [see link below], find the needed restriction enzyme sites that flank your desired 6kb sequence and clone the DNA into the vector. Be sure to check your sequence for any internal restriction sites. This may require you to do a partial digestion and recovery.
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Transglutaminases are a family of enzymes (EC 2.3.2.13) that catalyze the formation of a covalent bond between a free amine group (e.g., protein- or peptide-bound lysine) and the gamma-carboxamid group of protein- or peptide bound glutamine. Bonds formed by transglutaminase exhibit high resistance to proteolytic degradation.\n\nTransglutaminases were first described in 1957.[1]. The exact biochemical activity of transglutaminases was discovered in blood coagulation protein factor XIII in 1968.[2]\n\nPhysiological transglutaminases\nEight transglutaminases have been characterised.[3]\n\nName Activity Chromosome OMIM \nFactor XIII (fibrin-stabilizing factor) coagulaton 6p25-p24 Mendelian Inheritance in Man (OMIM) 134570 \nKeratinocyte transglutaminase (TGM1) skin 14q11.2 Mendelian Inheritance in Man (OMIM) 190195 \nTissue transglutaminase (TGM2) ubiquitous 20q11.2-q12 Mendelian Inheritance in Man (OMIM) 190196 \nEpidermal transglutaminase (TGM3) skin 20q12 Mendelian Inheritance in Man (OMIM) 600238 \nProstate transglutaminase (TGM4) prostate 3p22-p21.33 Mendelian Inheritance in Man (OMIM) 600585 \nTGM X or TGM5[4] skin 15q15.2 Mendelian Inheritance in Man (OMIM) 603805 \nTGM Y or TGM 6 unclear 20q11-15 not assigned \nTGM Z or TGM 7 testis, lung 15q15.2 Mendelian Inheritance in Man (OMIM) 606776 \n\nMechanism of action\nTransglutaminases form extensively cross linked generally insoluble protein polymers. These biological polymers are indispensable for the organism in order to create barriers and stable structures. Examples are blood clots (coagulation factor XIII) as well as skin and hair. The catalytic reaction is generally viewed as being irreversible and must be closely monitored through extensive control mechanisms.\n\nRole in disease\nDeficiency of factor XIII (a rare genetic condition) predisposes to hemorrhage; concentrated enzyme can be used to correct the abnormality and reduce bleeding risk.\n\nAntibodies to tissue transglutaminase are found in coeliac disease and may play a role in the small bowel damage in response to dietary gliadin that characterises this condition.\n\nRecent research indicates that sufferers from neurological diseases like Huntington's,[5] and Parkinson's[6] may have unusually high levels of one type of transglutaminase, tissue transglutaminase. It is hypothesized that tissue transglutaminase may be involved in the formation of the protein aggregates that causes Huntington's disease, although it is most likely not required.[7]\n\nIndustrial applications\nIndustrial transglutaminase is produced by Streptoverticillium mobaraense fermentation in commercial quantities and is used in a variety of processes, including the production of processed meat and fish products. It can be used as a binding agent to improve the texture of protein-rich foods such as surimi or ham.\n\nTransglutaminase can be used in these applications:\n\nBinding small chunks of meats into a big one ("portion control"), such as in sausages, hot dogs, restructured steaks \nImproving the texture of low-grade meat such as so-called "PSE meat" (pale, soft, and exudative meat; caused by stress and a rapid postmortem pH decline) \nMaking milk and yogurt creamier \nMaking noodles firmer \nBesides its "orthodox" uses, transglutaminase can be used to create some unusual foods. "Cold Set Bound Fish Kebabs" are made from alternating layers of salmon and cod which are "glued" together by transglutaminase. Wylie Dufresne, chef of New York's avant-garde restaurant WD-50, invented a "pasta" made by over 95% shrimps thanks to transglutaminase.
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rflp's are restriction fragment length polymorprhisms. Restriction fragments are obtained by taking a length of DNA and cutting it with restriction enzymes. Restriction enzymes are enzymes which cut DNA at a specific sequence and only that sequence. So the enzyme searches thru the DNA chain for a sequence that it can work on (there are different enzymes which work with different sequences) and cuts. If you take a chromosome and incubate it with your preferred enzyme the chromosome will be cut at specific sites wherever they appear in the sequence. This will give you a given number of fragments. Now each person's chromosomes are slightly different so if you took say your chromosome 1 and my chromosome 1 and incubated them both with a set restriction enzyme the majority of cuts would be done in the same place. But if I, for example, had a mutation in one of the sites preventing the enzyme from recognizing it i would have one long fragment where you have two. this is a polymorphism (a difference). Forensics Teams use this technique to look at DNA to find differences between individuals (you will have polymorphisms in common with both your parents but not quite the same but a higher resemblance to them than you would to me)\n\nhope thats semi clear - and good basic genetics book should help further e.g. Essential Genetics by Hartl and Jones
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That all depends on the enzyme. It could denature a protein involved, making the enzyme useless, or it could speed up the enzyme's activity.
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Introduction. This is a brief outline of the steps necessary to produce mice with a mutation targeted to a specific gene. These animals are referred to as "knock-out" mice or "gene targeted" mice. The broad outline of the experiment includes: 1) the investigator constructs a gene targeting vector containing a mutation in the target gene; 2) the targeting vector is introduced into ES cells; 3) ES cell clones which undergo homologous recombination with the targeting vector are identified; 4) ES cell-mouse chimeras are produced; 5) the chimeras breed and transmit the chromosome with the targeted gene to their offspring; 6) homozygous animals are produced from the mating of hemizygous chimera offspring; and 7) the phenotype resulting from the genetic mutation is characterized. Core personnel are available for consultation on all aspects of gene targeting research. Contact Thom Saunders with any questions or Meet the Staff.  The Transgenic Core prioritizes all requests for service on a "first-come, first-serve" basis. Your requests for electroporation, clone expansion, and blastocyst microinjection will be added to the Transgenic Core work queue in the order that they are received. \n2. Plan the experiment. What is the purpose of your experiment? Do you want to completely ablate gene expression in all tissues? Do you want to analyze tissue specific regulatory sequences? Do you want to produce a model for tissue-specific gene deletions? You will need to obtain or clone the structural gene that you want to mutate. The source of the DNA is important since published studies demonstrate genomic DNA isogenic to the embryonic stem cell line give higher gene targeting efficiencies in ES cells (see Key Gene Targeting Papers). While complete sequence information is not necessary, a detailed restriction map of the gene is an irreplaceable tool. During the planning stages the material transfer agreements for R1 ES cells and other reagents should be initiated so that they are ready in time. Once a genomic clone of the gene is in hand and restriction mapped, then it is time to... \n\n3. Clone and the Verify the Targeting Vector. The Core has cloning vectors for investigators to use in gene targeting research. They include pPNT, a basic targeting vector, pNZTK2, designed to insert the lacZ gene behind the promoter of the targeted gene, and pflox, which is designed to introduce loxP sites into the gene. A complete materials transfer agreement is required before they can be distributed. You will need to determine which 5' and 3' fragments of the gene will be used to construct the targeting vector. Generally speaking, a couple of Kb on either end should give a reasonable gene targeting frequency. You will also need to consider the methods and probes that you will use to identify ES cell clones that have undergone homologous recombination with the targeting vector. This means a Southern blot approach with probes outside of the targeting vector or a PCR based assay with primers outside of the targeting vector. Once the orientation of the fragments in the targeting vector are confirmed and the probes are tested then it is time to... \n\n4. Electroporate ES Cells and Select Recombinants. ES cells are a very sensitive reagent, and most often the weakest link in the process of producing gene targeted mice. Fastidious tissue culture technique is necessary to maintain the totipotent state of the cells during electroporation, selection, and expansion. In order to maximize successful outcomes, the Transgenic Core maintains ES cells and companion reagents that are quality tested by producing ES cell-mouse chimeras that transmit the ES cell genome to offspring. Investigators may choose to use the Core's Gene Targeting Service (we do all of the tissue culture work, you do all of the molecular biology), or to use space in the Core lab (rent space in our completely equipped laboratory for ES cell culture work), or to do th
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By the use of the enzyme DNA helicase
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"Currently, severe thalassemia is treated by blood transfusions, and a minority of patients are cured by bone marrow transplantation. Mouse models are proving to be useful in assessing the potential of gene therapy."\n\nhttp://www.ncbi.nlm.nih.gov/books/bv.fcgi?call=bv.View..ShowSection&rid=gnd.section.250
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maybe its bcoz of the genetic factor \n\nyoga is the best option \n\nit doesnot have side effects
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Not sure if this will help but try the link below\n\nhttp://www.answers.com/topic/gene
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If you are in Europe try Novozime (Netherland)\nIf you are in USA try Genencor \n\nTelomerase is an enzyme that take part in replication mechanism, especial in cancer cells (they do not lose a part of its DNA when the two strands are copied)
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maybe through receint genetic mutations that would enable them to produce enzymes that digest lactose in milk
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Restriction enzymes, also called restriction nucleases (EcoRI in this example) , surrounds the DNA molecule at the point it seeks(sequence GAATTC). It cuts one strand of the DNA double helix at one point and the second strand at a different, complementary point (between the G and the A base). The separated pieces have single stranded "sticky-ends," which allow the complementary pieces to combine. \n\nThe newly joined pieces are stabilized by DNA ligase.\n\nEcoRI, one of many restriction enzymes, is obtained from the bacteria Escherichia coli.
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figure out the efgfect of the enzyme. Or you can test DNA structure. Inhibitary enzymes have a blacked actionsite whereas activation enzymes have activation sites open to substrates
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Proc Natl Acad Sci U S A. 1990 November; 87(21): 8531–8535. \n\nHunter syndrome: isolation of an iduronate-2-sulfatase cDNA clone and analysis of patient DNA.\nP J Wilson, C P Morris, D S Anson, T Occhiodoro, J Bielicki, P R Clements, and J J Hopwood\n\nIduronate 2-sulfatase (IDS, EC 3.1.6.13) is required for the lysosomal degradation of heparan sulfate and dermatan sulfate. Mutations causing IDS deficiency in humans result in the lysosomal storage of these glycosaminoglycans and Hunter syndrome, an X chromosome-linked disease. We have isolated and sequenced a 2.3-kilobase cDNA clone coding for the entire sequence of human IDS. Analysis of the deduced 550-amino acid IDS precursor sequence indicates that IDS has a 25-amino acid amino-terminal signal sequence, followed by 8 amino acids that are removed from the proprotein. An internal proteolytic cleavage occurs to produce the mature IDS present in human liver shown to contain a 42-kDa polypeptide N-terminal to a 14-kDa polypeptide. The IDS sequence has strong sequence homology with other sulfatases (such as sea urchin arylsulfatase, human arylsulfatases A, B, and C, and human glucosamine 6-sulfatase), suggesting that the sulfatases comprise an evolutionarily related family of genes that arose by gene duplication and divergent evolution. The arylsulfatases have a greater homology with each other than with the non-arylsulfatases (IDS and glucosamine 6-sulfatase). The IDS cDNA detected RNA species of 5.7, 5.4, 2.1, and 1.4 kilobases in human placental RNA and revealed structural alterations and gross deletions of the IDS gene in many of the clinically severe Hunter syndrome patients studied.
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K.C.'s answer works if you have a selective media that will work. Also (or in addition to that) you may want to plate out colonies and then PCR out a unique sequence or two.\n\nThis wouldn't be definitive but if you design your primers right it may work, but also may be very time consuming.\n\nAlso you may want to try restriction digests to remap your gene. This would work well, if you have a good map.\n\nGood luck, what a bummer