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dc.creatorEnyeart, Peter J.en
dc.creatorMohr, Georgen
dc.creatorEllington, Andrew D.en
dc.creatorLambowitz, Alan M.en
dc.date.accessioned2014-12-15T17:10:53Zen
dc.date.available2014-12-15T17:10:53Zen
dc.date.issued2014-01-13en
dc.identifier.citationEnyeart, Peter J., Georg Mohr, Andrew D. Ellington, and Alan M. Lambowitz. “Biotechnological Applications of Mobile Group II Introns and Their Reverse Transcriptases: Gene Targeting, RNA-Seq, and Non-Coding RNA Analysis.” Mobile DNA 5, no. 1 (January 13, 2014): 2. doi:10.1186/1759-8753-5-2.en
dc.identifier.urihttp://hdl.handle.net/2152/27948en
dc.descriptionAll authors are with the Departments of Molecular Biosciences and Chemistry, Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX 78712, USAen
dc.description.abstractMobile group II introns are bacterial retrotransposons that combine the activities of an autocatalytic intron RNA (a ribozyme) and an intron-encoded reverse transcriptase to insert site-specifically into DNA. They recognize DNA target sites largely by base pairing of sequences within the intron RNA and achieve high DNA target specificity by using the ribozyme active site to couple correct base pairing to RNA-catalyzed intron integration. Algorithms have been developed to program the DNA target site specificity of several mobile group II introns, allowing them to be made into ‘targetrons.’ Targetrons function for gene targeting in a wide variety of bacteria and typically integrate at efficiencies high enough to be screened easily by colony PCR, without the need for selectable markers. Targetrons have found wide application in microbiological research, enabling gene targeting and genetic engineering of bacteria that had been intractable to other methods. Recently, a thermostable targetron has been developed for use in bacterial thermophiles, and new methods have been developed for using targetrons to position recombinase recognition sites, enabling large-scale genome-editing operations, such as deletions, inversions, insertions, and ‘cut-and-pastes’ (that is, translocation of large DNA segments), in a wide range of bacteria at high efficiency. Using targetrons in eukaryotes presents challenges due to the difficulties of nuclear localization and sub-optimal magnesium concentrations, although supplementation with magnesium can increase integration efficiency, and directed evolution is being employed to overcome these barriers. Finally, spurred by new methods for expressing group II intron reverse transcriptases that yield large amounts of highly active protein, thermostable group II intron reverse transcriptases from bacterial thermophiles are being used as research tools for a variety of applications, including qRT-PCR and next-generation RNA sequencing (RNA-seq). The high processivity and fidelity of group II intron reverse transcriptases along with their novel template-switching activity, which can directly link RNA-seq adaptor sequences to cDNAs during reverse transcription, open new approaches for RNA-seq and the identification and profiling of non-coding RNAs, with potentially wide applications in research and biotechnology.en
dc.description.sponsorshipen
dc.language.isoEnglishen
dc.publisherMobile DNA Journalen
dc.rightsAdministrative deposit of works to UT Digital Repository: This works author(s) is or was a University faculty member, student or staff member; this article is already available through open access at http://www.biomedcentral.com. The public license is specified as CC-BY: http://creativecommons.org/licenses/by/4.0/. The library makes the deposit as a matter of fair use (for scholarly, educational, and research purposes), and to preserve the work and further secure public access to the works of the University.en
dc.subjectGenome engineeringen
dc.subjectmetabolic engineeringen
dc.subjectnext-generation RNA sequencingen
dc.subjectribosymeen
dc.subjectsynthetic biologyen
dc.subjectsystems biologyen
dc.subjecttargetronen
dc.titleBiotechnological applications of mobile group II introns and their reverse transcriptases: gene targeting, RNA-seq, and non-coding RNA analysisen
dc.typeArticleen
dc.description.departmentMolecular Biosciencesen
dc.description.departmentChemistryen
dc.description.departmentInstitute for Cellular and Molecular Biologyen
dc.description.catalogingnotelambowitz@austin.utexas.eduen
dc.identifier.Filename1759-8753-5-2.pdfen
dc.identifier.doidoi:10.1186/1759-8753-5-2en


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