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Videos de Conceptos Relacionados

Conservative Site-specific Recombination and Phase Variation02:53

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
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DNA-only Transposons02:57

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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
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Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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LTR Retrotransposons03:08

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LTR retrotransposons are class I transposable elements with long terminal repeats flanking an internal coding region. These elements are less abundant in mammals compared to other class I transposable elements. About 8 percent of human genomic DNA comprises LTR retrotransposons. Some of the common examples of LTR retrotransposons are Ty elements in yeast and Copia elements in Drosophila.
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Non-LTR Retrotransposons03:18

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As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
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Introduction to Nuclear Reprogramming01:14

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Nuclear reprogramming is the process of switching gene expression of one cell type to that of another cell type, usually from a differentiated cell state to an undifferentiated cell state. Differentiation occurs during processes such as development and morphogenesis, tissue regeneration, and malignancy. Cells can also be artificially induced to reprogram their gene expression by techniques such as nuclear transfer, induced pluripotency, and cell fusion. Such techniques have many applications in...
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Reprogramación de la actividad de retrotransposón específica del sitio a nuevos sitios de ADN

Christopher W Fell1,2,3,4, Lukas Villiger4, Justin Lim4

  • 1Department of Medicine, Division of Engineering in Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.

Nature
|April 9, 2025
PubMed
Resumen

Los investigadores diseñaron un nuevo sistema llamado STITCHR para la edición precisa y sin cicatrices del genoma. Esta herramienta basada en retroelementos permite la inserción eficiente de material genético en ubicaciones específicas tanto en células que se dividen como en células que no se dividen.

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Área de la Ciencia:

  • Genómica y Biología Molecular
  • Biología del retrotransposón
  • Tecnologías de edición de genes

Sus antecedentes:

  • Las retrotransposones de repetición terminal no larga (no LTR) son los impulsores clave de la evolución del genoma eucariota.
  • Estos elementos genéticos móviles a menudo se integran en regiones genómicas repetitivas específicas.
  • Los mecanismos de orientación precisos y las limitaciones de los retrotransposones siguen siendo incompletamente entendidos.

Objetivo del estudio:

  • Descubrir y caracterizar nuevas familias de retrotransposones específicas del sitio.
  • Investigar las preferencias de inserción y el potencial de retargeting de los retrotransposones.
  • Para diseñar una nueva plataforma para la integración genómica precisa y sin cicatrices.

Principales métodos:

  • Utilizó una tubería computacional para identificar nuevas familias de retrotransposones.
  • Se realizó un perfilado bioquímico y celular de los miembros del retrotransposón identificados.
  • Diseñó un sistema de fusión retrotransposón-CRISPR (STITCHR) para la inserción dirigida.

Principales resultados:

  • Se descubrieron nuevas familias de retrotransposones específicas del sitio con nuevas preferencias de inserción.
  • Con éxito redirigido un retrotransposon R2 (R2Tg) para la inserción sin cicatrices de las cargas útiles.
  • Desarrolló STITCHR, lo que permite una instalación eficiente y sin cicatrices de ediciones de hasta 12.7 kb, reemplazo de genes y uso de plantillas de ARN.

Conclusiones:

  • STITCHR representa una plataforma versátil para la ingeniería del genoma sin cicatrices y programable.
  • El sistema demuestra el potencial de aplicaciones tanto en la investigación como en el ámbito terapéutico.
  • Este enfoque aprovecha la prevalencia natural de retrotransposones no LTR para la edición genética avanzada.