Implementación de tecnología Crispr/Cas9 aplicada en Caenorhabditis Elegans como Posible opción terapéutica para la enfermedad de Alzheimer revisión documental
La enfermedad de Alzheimer (EA) es la demencia más común, caracterizada por la disminución gradual y progresiva de la memoria y la cognición, una raíz de distintos mecanismos, siendo la formación de proteína β-amiloide (Aβ) el proceso más conocido. En Colombia no existen datos epidemiológicos actual...
- Autores:
-
Neira Mora, Daniel Gustavo
- Tipo de recurso:
- Trabajo de grado de pregrado
- Fecha de publicación:
- 2020
- Institución:
- Colegio Mayor de Cundinamarca
- Repositorio:
- Repositorio Colegio Mayor de Cundinamarca
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.unicolmayor.edu.co:unicolmayor/68
- Acceso en línea:
- https://repositorio.unicolmayor.edu.co/handle/unicolmayor/68
- Palabra clave:
- Caenorhabditis Elegans
Alzheimer
Aβ
Tau
Crispr/Cas9
- Rights
- closedAccess
- License
- Derechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2020
| id |
UCOLMAYOR2_c69eb35d6f3ef59759b697a04f2fe10f |
|---|---|
| oai_identifier_str |
oai:repositorio.unicolmayor.edu.co:unicolmayor/68 |
| network_acronym_str |
UCOLMAYOR2 |
| network_name_str |
Repositorio Colegio Mayor de Cundinamarca |
| repository_id_str |
|
| dc.title.spa.fl_str_mv |
Implementación de tecnología Crispr/Cas9 aplicada en Caenorhabditis Elegans como Posible opción terapéutica para la enfermedad de Alzheimer revisión documental |
| title |
Implementación de tecnología Crispr/Cas9 aplicada en Caenorhabditis Elegans como Posible opción terapéutica para la enfermedad de Alzheimer revisión documental |
| spellingShingle |
Implementación de tecnología Crispr/Cas9 aplicada en Caenorhabditis Elegans como Posible opción terapéutica para la enfermedad de Alzheimer revisión documental Caenorhabditis Elegans Alzheimer Aβ Tau Crispr/Cas9 |
| title_short |
Implementación de tecnología Crispr/Cas9 aplicada en Caenorhabditis Elegans como Posible opción terapéutica para la enfermedad de Alzheimer revisión documental |
| title_full |
Implementación de tecnología Crispr/Cas9 aplicada en Caenorhabditis Elegans como Posible opción terapéutica para la enfermedad de Alzheimer revisión documental |
| title_fullStr |
Implementación de tecnología Crispr/Cas9 aplicada en Caenorhabditis Elegans como Posible opción terapéutica para la enfermedad de Alzheimer revisión documental |
| title_full_unstemmed |
Implementación de tecnología Crispr/Cas9 aplicada en Caenorhabditis Elegans como Posible opción terapéutica para la enfermedad de Alzheimer revisión documental |
| title_sort |
Implementación de tecnología Crispr/Cas9 aplicada en Caenorhabditis Elegans como Posible opción terapéutica para la enfermedad de Alzheimer revisión documental |
| dc.creator.fl_str_mv |
Neira Mora, Daniel Gustavo |
| dc.contributor.advisor.none.fl_str_mv |
Sánchez Mora, Ruth Mélida Rodríguez Panduro, Mauricio Humberto Salazar Buitrago, Nelson Arturo |
| dc.contributor.author.none.fl_str_mv |
Neira Mora, Daniel Gustavo |
| dc.subject.lemb.none.fl_str_mv |
Caenorhabditis Elegans Alzheimer Aβ Tau Crispr/Cas9 |
| topic |
Caenorhabditis Elegans Alzheimer Aβ Tau Crispr/Cas9 |
| description |
La enfermedad de Alzheimer (EA) es la demencia más común, caracterizada por la disminución gradual y progresiva de la memoria y la cognición, una raíz de distintos mecanismos, siendo la formación de proteína β-amiloide (Aβ) el proceso más conocido. En Colombia no existen datos epidemiológicos actualizados, por lo que resulta imposible acceder a un dato preciso de esta enfermedad. Tras 25 años no se ha obtenido un tratamiento eficaz y dilucidar los mecanismos de la enfermedad, ha sido una tarea difícil dada la complejidad del cerebro humano, por esta razón, el modelado de la EA en animales con estructuras nerviosas conocidas ofrece grandes avances. Caenorhabditis elegans (C. elegans) es un nematodo de vida libre ampliamente utilizado como modelo neurobiológico, dado que su genoma contiene un 60% de similitud con el genoma humano. CRISPR/Cas9 ha sido la herramienta de edición genética más revolucionaria desde su descubrimiento en Arqueas. El sistema CRISPR tipo II (Cas9) es por mucho, la tecnología mejor desarrollada para editar cualquier gen, esto lo convierte en una alternativa con potencial terapéutico en EA. Conociendo las hipótesis de la cascada amiloide, Presenilinas y enfoque de matriz APP como principales hipótesis de EA, es posible determinar el papel en la progresión de EA que tiene genes como APP, PSEN, ApoE, SORL1, ABCA7 entre otros. Esto permite modelar Aβ, APP y Tau en cepas de C. elegans, para identificar estrategias terapéuticas de CRISPR/Cas9; Ya sea en el aclaramiento de Aβ o corrigiendo mutaciones ya conocidas implicadas en EA. |
| publishDate |
2020 |
| dc.date.issued.none.fl_str_mv |
2020-06-02 |
| dc.date.accessioned.none.fl_str_mv |
2021-05-20T13:36:46Z |
| dc.date.available.none.fl_str_mv |
2021-05-20T13:36:46Z |
| dc.type.spa.fl_str_mv |
Trabajo de grado - Pregrado |
| dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_7a1f |
| dc.type.coarversion.spa.fl_str_mv |
http://purl.org/coar/version/c_b1a7d7d4d402bcce |
| dc.type.content.spa.fl_str_mv |
Text |
| dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/bachelorThesis |
| dc.type.redcol.spa.fl_str_mv |
https://purl.org/redcol/resource_type/TP |
| dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/draft |
| format |
http://purl.org/coar/resource_type/c_7a1f |
| status_str |
draft |
| dc.identifier.uri.none.fl_str_mv |
https://repositorio.unicolmayor.edu.co/handle/unicolmayor/68 |
| url |
https://repositorio.unicolmayor.edu.co/handle/unicolmayor/68 |
| dc.language.iso.spa.fl_str_mv |
spa |
| language |
spa |
| dc.relation.references.spa.fl_str_mv |
Byerly L, Cassada RC, Russell RL. The life cycle of the nematode Caenorhabditis elegans. I. Wild-type growth and reproduction. Dev Biol 1976 Jul 01,;51(1):23-33. Kenyon C. The Nematode Caenorhabditis elegans. Science 1988;240(4858):1448-1453. Blaxter M. Caenorhabditis elegans is a nematode. Science 1998 Dec 11,;282(5396):2041-2046. Culetto E, Sattelle DB. A role for Caenorhabditis elegans in understanding the function and interactions of human disease genes. Hum Mol Genet 2000 Apr 12,;9(6):869-877. Hodgkin J. Caenorhabditis Elegans. Encyclopedia of Genetics 2001:251-256. Chisholm AD, Jin Y. Neuronal differentiation in C. elegans. Current Opinion in Cell Biology 2005;17(6):682-689. Schafer WR. Deciphering the neural and molecular mechanisms of C. elegans behavior. Curr Biol 2005 Sep 06,;15(17):723. Miyasaka T, Ding Z, Gengyo-Ando K, Oue M, Yamaguchi H, Mitani S, et al. Progressive neurodegeneration in C. elegans model of tauopathy. Neurobiology of Disease 2005;20(2):372-383. Coghlan A. Nematode genome evolution: WormBook; 2005. Savel J, Clostre F. [A Nematode Nobel Prize: Caenorhabditis elegans]. Ann Pharm Fr 2006 Sep;64(5):291-307. Fay D. Genetic mapping and manipulation: Chapter 1-Introduction and basics. WormBook : the online review of C. elegans biology; 2006:1-12. Fay D. Genetic mapping and manipulation: chapter 9--Synthetic and enhancer mutations. WormBook 2006 Feb 17,:1-7. Praitis V. Creation of transgenic lines using microparticle bombardment methods. Methods Mol Biol 2006;351:93-107. Kaletta T, Hengartner MO. Finding function in novel targets: C. elegans as a model organism. Nat Rev Drug Discov 2006 05;5(5):387-398. Gregorio. P, Sanchez. J. DEMENCIA. Sociedad Española de Geriatría y Gerontología, editor. Tratado de geriatría para residentes Madrid: International Marketing & Communication, S.A.; 2006. p. 173-188. Holmes C. Dementia. Medicine 2008;36(9):467-470 Lee M, Chodosh J. Dementia and Life Expectancy: What Do We Know? Journal of the American Medical Directors Association 2009;10(7):466-471 Chen J, Lin K, Chen Y. Risk Factors for Dementia. Journal of the Formosan Medical Association 2009;108(10):754-764. Ramirez-Bermudez J. Alzheimer's Disease: Critical Notes on the History of a Medical Concept. Archives of Medical Research 2012;43(8):595- 599. Gazes Y, Soldan A, Stern Y. Alzheimer's Disease. Encyclopedia of Human Behavior (Second Edition) 2012:108-115. Walsh DM, Teplow DB. Chapter 4 - Alzheimer's Disease and the Amyloid β-Protein. Progress in Molecular Biology and Translational Science 2012;107:101-124. Castellani RJ, Moreira PI, Zhu X, Perry G. Alzheimer's Disease: Pathology and Pathogenesis. Reference Module in Biomedical Sciences 2014. Peter J, Klöppel S. Alzheimer's Disease. Brain Mapping 2015:647-651. Merino EN, Sendin MAC, Osorio JAV. Enfermedad de Alzheimer. Medicine - Programa de Formación Médica Continuada Acreditado 2015;11(72):4306-4315. Braak H, Del Tredici-Braak K. Alzheimer's Disease, Neural Basis of. International Encyclopedia of the Social & Behavioral Sciences (Second Edition) 2015:591-596. Reitz C. Genetic diagnosis and prognosis of Alzheimer's disease: challenges and opportunities. Expert Review of Molecular Diagnostics 2015 Mar;15(3):339-348. Tekeuchi Y. ALZHEIMER UN PROBLEMA DE SALUD PÚBLICA EN COLOMBIA. 2015; Available at: https://www.icesi.edu.co/unicesi/todas-las- noticias/2241-alzheimer-un-problema-de-salud-publica-en-colombia. Accessed Mar 18, 2020. Arendt T, Stieler JT, Holzer M. Tau and tauopathies. Brain Research Bulletin 2016;126:238-292. Selkoe DJ, Hardy J. The amyloid hypothesis of Alzheimer's disease at 25 years. EMBO molecular medicine 2016 Jun;8(6):595-608. Kanekiyo T, Bu G. Chapter 6 - Apolipoprotein E and Amyloid-βIndependent Mechanisms in Alzheimer’s Disease. Genes, Environment and Alzheimer's Disease 2016:171-196. Xue-shan Z, juan P, Qi W, Zhong R, Li-hong P, Zhi-han T, et al. Imbalanced cholesterol metabolism in Alzheimer's disease. Clinica Chimica Acta 2016;456:107-114. Hung ASM, Liang Y, Chow TCH, Tang HC, Wu SLY, Wai MSM, et al. Mutated tau, amyloid and neuroinflammation in Alzheimer disease—A brief review. Progress in Histochemistry and Cytochemistry 2016;51(1):1-8. De Strooper B, Karran E. The Cellular Phase of Alzheimer's Disease. Cell 2016 Feb 11,;164(4):603-615. Luczkowski M. “No screams and cries will convince us that white is white and black is black”, an ode to the defenders of amyloid cascade hypothesis of Alzheimer's disease. Coordination Chemistry Reviews 2016;327-328:35-42 Calsolaro V, Edison P. Neuroinflammation in Alzheimer's disease: Current evidence and future directions. Alzheimer's & Dementia 2016;12(6):719-732. Hooli B, Tanzi RE. Chapter 2 - The Genetic Basis of Alzheimer’s Disease. Developing Therapeutics for Alzheimer's Disease 2016 :23-37. Hooli B, Tanzi RE. Chapter 34 - The Genetic Basis of Alzheimer's Disease: Findings From Genome-Wide Studies. Genomics, Circuits, and Pathways in Clinical Neuropsychiatry 2016:547-571 Tosto G, Reitz C. Genomics of Alzheimer's disease: Value of highthroughput genomic technologies to dissect its etiology. Molecular and Cellular Probes 2016;30(6):397-403. Mohamed T, Shakeri A, Rao PPN. Amyloid cascade in Alzheimer's disease: Recent advances in medicinal chemistry. European Journal of Medicinal Chemistry 2016;113:258-272. Sterner RM, Takahashi PY, Yu Ballard AC. Active Vaccines for Alzheimer Disease Treatment. Journal of the American Medical Directors Association 2016;17(9):862.e1-862.e15. López Casillas F, López Casillas F. CRISPR, el sueño divino hecho realidad. Revista de la Facultad de Medicina (México) 2016 08/;58(4):55-60. Kanchiswamy CN, Maffei M, Malnoy M, Velasco R, Kim J. Fine-Tuning Next-Generation Genome Editing Tools. Trends in Biotechnology 2016;34(7):562-574. Maeder ML, Gersbach CA. Genome-editing Technologies for Gene and Cell Therapy. Molecular Therapy 2016;24(3):430-446. Prabhu V, Xu H. Endonuclease mediated genome editing in drug discovery and development: promises and challenges. Drug Discovery Today: Technologies 2016;21-22:17-25. Lander E. The Heroes of CRISPR. Cell 2016;164(1):18-28. Mojica FJM, Montoliu L. On the Origin of CRISPR-Cas Technology: From Prokaryotes to Mammals. Trends in Microbiology 2016;24(10):811-820. Golubov A. Chapter 6 - CRISPR: Bacteria Immune System. Genome Stability 2016:87-98. Han W, She Q. Chapter One - CRISPR History: Discovery, Characterization, and Prosperity. Progress in Molecular Biology and Translational Science 2017;152:1-21 Ede DR, Farhang N, Stover JD, Bowles RD. 4.32 Gene Editing Tools. Comprehensive Biomaterials II 2017:589-599. Pelletier S. CRISPR–Cas9. Encyclopedia of Microbiology (Fourth Edition) 2017:772-782. Lemay M, Horvath P, Moineau S. The CRISPR-Cas app goes viral. Current Opinion in Microbiology 2017;37:103-109. Hille F, Richter H, Wong SP, Bratovič M, Ressel S, Charpentier E. The Biology of CRISPR-Cas: Backward and Forward. Cell 2018;172(6):1239-1259 Singh V, Braddick D, Dhar PK. Exploring the potential of genome editing CRISPR-Cas9 technology. Gene 2017;599:1-18. Ramirez JC. Chapter Six - Gene Editing and CRISPR Therapeutics: Strategies Taught by Cell and Gene Therapy. Progress in Molecular Biology and Translational Science 2017;152:115-130. Martinez-Lage M, Torres-Ruiz R, Rodriguez-Perales S. Chapter Two - CRISPR/Cas9 Technology: Applications and Human Disease Modeling. Progress in Molecular Biology and Translational Science 2017;152:23-48. Powell SK, Gregory J, Akbarian S, Brennand KJ. Application of CRISPR/Cas9 to the study of brain development and neuropsychiatric disease. Molecular and Cellular Neuroscience 2017;82:157-166. Shrock E, Güell M. Chapter Six - CRISPR in Animals and Animal Models. Progress in Molecular Biology and Translational Science 2017;152:95-114. de Lecuona I, Casado M, Marfany G, Lopez Baroni M, Escarrabill M. Gene Editing in Humans: Towards a Global and Inclusive Debate for Responsible Research. Yale J Biol Med 2017 12;90(4):673-681. Cribbs AP, Perera SMW. Science and Bioethics of CRISPR-Cas9 Gene Editing: An Analysis Towards Separating Facts and Fiction. Yale J Biol Med 2017 12;90(4):625-634. Griffin EF, Caldwell KA, Caldwell GA. Genetic and Pharmacological Discovery for Alzheimer's Disease Using Caenorhabditis elegans. ACS Chem Neurosci 2017 12 20,;8(12):2596-2606. Pir GJ, Choudhary B, Mandelkow E. Caenorhabditis elegans models of tauopathy. FASEB J 2017 12;31(12):5137-5148 Ortiz-Virumbrales M, Moreno CL, Kruglikov I, Marazuela P, Sproul A, Jacob S, et al. CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer's PSEN2 N141I neurons. Acta Neuropathol Commun 2017 10 27,;5(1):77. Shao W, Peng D, Wang X. Genetics of Alzheimer’s disease: From pathogenesis to clinical usage. Journal of Clinical Neuroscience 2017;45:1-8. Carmona S, Hardy J, Guerreiro R. Chapter 26 - The genetic landscape of Alzheimer disease. Handbook of Clinical Neurology 2018;148:395-408. Hunter S, Smailagic N, Brayne C. Aβ and the dementia syndrome: Simple versus complex perspectives. European Journal of Clinical Investigation 2018;48(12):e13025. Kim JH. Genetics of Alzheimer's Disease. Dement Neurocogn Disord 2018;17(4):131-136. McDiarmid TA, Au V, Loewen AD, Liang J, Mizumoto K, Moerman DG, et al. CRISPR-Cas9 human gene replacement and phenomic characterization in Caenorhabditis elegans to understand the functional conservation of human genes and decipher variants of uncertain significance. Dis Model Mech 2018 11 26,;11(12). Rohn TT, Kim N, Isho NF, Mack JM. The Potential of CRISPR/Cas9 Gene Editing as a Treatment Strategy for Alzheimer’s Disease. J Alzheimers Dis Parkinsonism 2018;8(3) Rahman S, Datta M, Kim J, Jan AT. CRISPR/Cas: An intriguing genomic editing tool with prospects in treating neurodegenerative diseases. Seminars in Cell & Developmental Biology 2019;96:22-31. Roshni Mishra, Leya Joykutty, Kepa Oyarbide American Heritage School, Plantation, FL. Expression of anti-neurodegeneration genes in mutant Caenorhabditis elegans using CRISPR-Cas9 improves behavior associated with Alzheimer’s Disease 2019. Ins. Europeo de Bioinformática, Ins. Wellcome Trust Sanger, Ins. Ontario para la Investigación del Cáncer. WormBase. Available at: https://wormbase.org, 2020. |
| dc.rights.spa.fl_str_mv |
Derechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2020 |
| dc.rights.uri.spa.fl_str_mv |
https://creativecommons.org/licenses/by-nc-sa/4.0/ |
| dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/closedAccess |
| dc.rights.creativecommons.spa.fl_str_mv |
Atribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0) |
| dc.rights.coar.spa.fl_str_mv |
http://purl.org/coar/access_right/c_14cb |
| rights_invalid_str_mv |
Derechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2020 https://creativecommons.org/licenses/by-nc-sa/4.0/ Atribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0) http://purl.org/coar/access_right/c_14cb |
| eu_rights_str_mv |
closedAccess |
| dc.format.extent.spa.fl_str_mv |
106p. |
| dc.format.mimetype.spa.fl_str_mv |
application/pdf |
| dc.publisher.spa.fl_str_mv |
Universidad Colegio Mayor de Cundinamarca |
| dc.publisher.faculty.spa.fl_str_mv |
Facultad de Ciencias de la Salud |
| dc.publisher.place.spa.fl_str_mv |
Bogotá D.C |
| dc.publisher.program.spa.fl_str_mv |
Bacteriología y Laboratorio Clínico |
| institution |
Colegio Mayor de Cundinamarca |
| bitstream.url.fl_str_mv |
https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/4/license.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/11/Cartas%20derechos%20de%20autor.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/13/INFORME%20FINAL%202020.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/12/Cartas%20derechos%20de%20autor.pdf.jpg https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/14/INFORME%20FINAL%202020.pdf.jpg https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/15/INFORME%20FINAL%202020.pdf https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/16/Cartas%20derechos%20de%20autor.pdf |
| bitstream.checksum.fl_str_mv |
2f9959eaf5b71fae44bbf9ec84150c7a 269ba7e629f8fadbe92d99e25fe0bd8a 8cc25273cd05f20f9985f9e8bf21442d 2f830c4815c93690e5d7f0f1564fa2f3 bb2e472d25e8f261f9caa11497735dd3 36bb7472d24369b848ccfd6fab4d068e bfeb2b90137f3c02a4ae30abaf872b85 |
| bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 MD5 MD5 |
| repository.name.fl_str_mv |
Biblioteca Digital Unicolmayor |
| repository.mail.fl_str_mv |
repositorio@unicolmayor.edu.co |
| _version_ |
1812210064258236416 |
| spelling |
Sánchez Mora, Ruth Mélidaefe8f5fbfdf7ec7c24ba850634bfbb9aRodríguez Panduro, Mauricio Humberto5a3aa353075aff39d48a3977c88cdb07Salazar Buitrago, Nelson Arturoe7b4c33854f42cc7b4ceb4e7c53977e2Neira Mora, Daniel Gustavoaa908a27f046f6685bc644fc6f2c1eb32021-05-20T13:36:46Z2021-05-20T13:36:46Z2020-06-02https://repositorio.unicolmayor.edu.co/handle/unicolmayor/68La enfermedad de Alzheimer (EA) es la demencia más común, caracterizada por la disminución gradual y progresiva de la memoria y la cognición, una raíz de distintos mecanismos, siendo la formación de proteína β-amiloide (Aβ) el proceso más conocido. En Colombia no existen datos epidemiológicos actualizados, por lo que resulta imposible acceder a un dato preciso de esta enfermedad. Tras 25 años no se ha obtenido un tratamiento eficaz y dilucidar los mecanismos de la enfermedad, ha sido una tarea difícil dada la complejidad del cerebro humano, por esta razón, el modelado de la EA en animales con estructuras nerviosas conocidas ofrece grandes avances. Caenorhabditis elegans (C. elegans) es un nematodo de vida libre ampliamente utilizado como modelo neurobiológico, dado que su genoma contiene un 60% de similitud con el genoma humano. CRISPR/Cas9 ha sido la herramienta de edición genética más revolucionaria desde su descubrimiento en Arqueas. El sistema CRISPR tipo II (Cas9) es por mucho, la tecnología mejor desarrollada para editar cualquier gen, esto lo convierte en una alternativa con potencial terapéutico en EA. Conociendo las hipótesis de la cascada amiloide, Presenilinas y enfoque de matriz APP como principales hipótesis de EA, es posible determinar el papel en la progresión de EA que tiene genes como APP, PSEN, ApoE, SORL1, ABCA7 entre otros. Esto permite modelar Aβ, APP y Tau en cepas de C. elegans, para identificar estrategias terapéuticas de CRISPR/Cas9; Ya sea en el aclaramiento de Aβ o corrigiendo mutaciones ya conocidas implicadas en EA.Alzheimer's disease (AD) is the most common dementia, characterized by the gradual and progressive decrease in memory and cognition, a root of the loss of neurons due to the action of different mechanisms, the formation of β-amyloid protein (Aβ) being the mechanism better known. In Colombia there are no updated epidemiological data, making it impossible to access accurate data on this disease. After 25 years, an effective treatment has not been obtained and elucidating the mechanisms of the disease, it has been a difficult task, due to the complexity of the human brain, for this reason, the modeling of AD in animals with known nerve structures offers great advances. Caenorhabditis elegans (C. elegans) is a nematode widely used as a neurobiological model, since its genome contains 60% similarity to the human genome. CRISPR / Cas9 has been the most revolutionary genetic editing tool since its discovery in Arqueas. The CRISPR type II system (Cas9) is by far the best developed technology to edit any gene, this makes it an alternative with therapeutic potential in AD. Knowing the hypotheses of the amyloid cascade, Presenilins and APP matrix approach as the main hypotheses of EA, it is possible to determine the role in the progression of EA that has genes such as APP, PSEN, ApoE, SORL1, ABCA7 among others. This allows to model Aβ, APP and Tau in strains of C. elegans, to identify therapeutic strategies of CRISPR / Cas9; Either in Aβ clearance or by correcting known mutations involved in EA.1. INTRODUCCIÓN 13 2. ANTECEDENTES 15 3. MARCO TEÓRICO 18 3.1 Caenorhabditis elegans 18 3.1.1 Ciclo de vida y desarrollo 19 3.1.2 Utilidad de C. elegans en investigación 21 3.2 Demencias y Enfermedad de Alzheimer 23 3.2.1 Etiología de las demencias 23 3.2.2 Clasificación de las demencias 24 3.3 Enfermedad de Alzheimer 25 3.3.1 Fisiopatología de la EA 27 3.3.2 Fase bioquímica de la EA 28 3.3.3 Fase celular de EA 35 3.4 Edición genética 38 5.1 CRISPR/Cas9 44 5.1.1 Estructura del Operón Cas y la matriz CRISPR 46 5.1.2 CRISPR TIPO II (Cas9) 47 5.2 Potencial de CRISPR/Cas9 en tratamiento de EA. 48 5.3 Hipótesis de EA 50 5.3.1 Hipótesis de la cascada amiloide 50 5.3.2 Hipótesis de la presenilina 51 5.3.3 Enfoque de la matriz APP 51 5.4 Genes implicados en la EA 52 5.5 Modelado de la EA en C. elegans y genes implicados 54 5.5.1 Modelado de Aβ en musculo 55 5.5.2 Modelado de Tau 56 5.6 Terapéutica usando CRISPR/CAS9 en EA Y C. elegans 60 5.6.1 Aclaramiento de proteínas responsables de EA en C. elegans 60 5.6.2 Corrección de mutaciones responsables de EA en C. elegans 62 6. DISCUSIÓN 64 7. CONCLUSIONES 68 8. BIBLIOGRAFÍA 69PregradoBacteriólogo(a) y Laboratorista Clínico106p.application/pdfspaUniversidad Colegio Mayor de CundinamarcaFacultad de Ciencias de la SaludBogotá D.CBacteriología y Laboratorio ClínicoDerechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2020https://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/closedAccessAtribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0)http://purl.org/coar/access_right/c_14cbImplementación de tecnología Crispr/Cas9 aplicada en Caenorhabditis Elegans como Posible opción terapéutica para la enfermedad de Alzheimer revisión documentalTrabajo de grado - Pregradohttp://purl.org/coar/resource_type/c_7a1fhttp://purl.org/coar/version/c_b1a7d7d4d402bcceTextinfo:eu-repo/semantics/bachelorThesishttps://purl.org/redcol/resource_type/TPinfo:eu-repo/semantics/draftByerly L, Cassada RC, Russell RL. The life cycle of the nematode Caenorhabditis elegans. I. Wild-type growth and reproduction. Dev Biol 1976 Jul 01,;51(1):23-33.Kenyon C. The Nematode Caenorhabditis elegans. Science 1988;240(4858):1448-1453.Blaxter M. Caenorhabditis elegans is a nematode. Science 1998 Dec 11,;282(5396):2041-2046.Culetto E, Sattelle DB. A role for Caenorhabditis elegans in understanding the function and interactions of human disease genes. Hum Mol Genet 2000 Apr 12,;9(6):869-877.Hodgkin J. Caenorhabditis Elegans. Encyclopedia of Genetics 2001:251-256.Chisholm AD, Jin Y. Neuronal differentiation in C. elegans. Current Opinion in Cell Biology 2005;17(6):682-689.Schafer WR. Deciphering the neural and molecular mechanisms of C. elegans behavior. Curr Biol 2005 Sep 06,;15(17):723.Miyasaka T, Ding Z, Gengyo-Ando K, Oue M, Yamaguchi H, Mitani S, et al. Progressive neurodegeneration in C. elegans model of tauopathy. Neurobiology of Disease 2005;20(2):372-383.Coghlan A. Nematode genome evolution: WormBook; 2005.Savel J, Clostre F. [A Nematode Nobel Prize: Caenorhabditis elegans]. Ann Pharm Fr 2006 Sep;64(5):291-307.Fay D. Genetic mapping and manipulation: Chapter 1-Introduction and basics. WormBook : the online review of C. elegans biology; 2006:1-12.Fay D. Genetic mapping and manipulation: chapter 9--Synthetic and enhancer mutations. WormBook 2006 Feb 17,:1-7.Praitis V. Creation of transgenic lines using microparticle bombardment methods. Methods Mol Biol 2006;351:93-107.Kaletta T, Hengartner MO. Finding function in novel targets: C. elegans as a model organism. Nat Rev Drug Discov 2006 05;5(5):387-398.Gregorio. P, Sanchez. J. DEMENCIA. Sociedad Española de Geriatría y Gerontología, editor. Tratado de geriatría para residentes Madrid: International Marketing & Communication, S.A.; 2006. p. 173-188.Holmes C. Dementia. Medicine 2008;36(9):467-470Lee M, Chodosh J. Dementia and Life Expectancy: What Do We Know? Journal of the American Medical Directors Association 2009;10(7):466-471Chen J, Lin K, Chen Y. Risk Factors for Dementia. Journal of the Formosan Medical Association 2009;108(10):754-764.Ramirez-Bermudez J. Alzheimer's Disease: Critical Notes on the History of a Medical Concept. Archives of Medical Research 2012;43(8):595- 599.Gazes Y, Soldan A, Stern Y. Alzheimer's Disease. Encyclopedia of Human Behavior (Second Edition) 2012:108-115.Walsh DM, Teplow DB. Chapter 4 - Alzheimer's Disease and the Amyloid β-Protein. Progress in Molecular Biology and Translational Science 2012;107:101-124.Castellani RJ, Moreira PI, Zhu X, Perry G. Alzheimer's Disease: Pathology and Pathogenesis. Reference Module in Biomedical Sciences 2014.Peter J, Klöppel S. Alzheimer's Disease. Brain Mapping 2015:647-651.Merino EN, Sendin MAC, Osorio JAV. Enfermedad de Alzheimer. Medicine - Programa de Formación Médica Continuada Acreditado 2015;11(72):4306-4315.Braak H, Del Tredici-Braak K. Alzheimer's Disease, Neural Basis of. International Encyclopedia of the Social & Behavioral Sciences (Second Edition) 2015:591-596.Reitz C. Genetic diagnosis and prognosis of Alzheimer's disease: challenges and opportunities. Expert Review of Molecular Diagnostics 2015 Mar;15(3):339-348.Tekeuchi Y. ALZHEIMER UN PROBLEMA DE SALUD PÚBLICA EN COLOMBIA. 2015; Available at: https://www.icesi.edu.co/unicesi/todas-las- noticias/2241-alzheimer-un-problema-de-salud-publica-en-colombia. Accessed Mar 18, 2020.Arendt T, Stieler JT, Holzer M. Tau and tauopathies. Brain Research Bulletin 2016;126:238-292.Selkoe DJ, Hardy J. The amyloid hypothesis of Alzheimer's disease at 25 years. EMBO molecular medicine 2016 Jun;8(6):595-608.Kanekiyo T, Bu G. Chapter 6 - Apolipoprotein E and Amyloid-βIndependent Mechanisms in Alzheimer’s Disease. Genes, Environment and Alzheimer's Disease 2016:171-196.Xue-shan Z, juan P, Qi W, Zhong R, Li-hong P, Zhi-han T, et al. Imbalanced cholesterol metabolism in Alzheimer's disease. Clinica Chimica Acta 2016;456:107-114.Hung ASM, Liang Y, Chow TCH, Tang HC, Wu SLY, Wai MSM, et al. Mutated tau, amyloid and neuroinflammation in Alzheimer disease—A brief review. Progress in Histochemistry and Cytochemistry 2016;51(1):1-8.De Strooper B, Karran E. The Cellular Phase of Alzheimer's Disease. Cell 2016 Feb 11,;164(4):603-615.Luczkowski M. “No screams and cries will convince us that white is white and black is black”, an ode to the defenders of amyloid cascade hypothesis of Alzheimer's disease. Coordination Chemistry Reviews 2016;327-328:35-42Calsolaro V, Edison P. Neuroinflammation in Alzheimer's disease: Current evidence and future directions. Alzheimer's & Dementia 2016;12(6):719-732.Hooli B, Tanzi RE. Chapter 2 - The Genetic Basis of Alzheimer’s Disease. Developing Therapeutics for Alzheimer's Disease 2016 :23-37.Hooli B, Tanzi RE. Chapter 34 - The Genetic Basis of Alzheimer's Disease: Findings From Genome-Wide Studies. Genomics, Circuits, and Pathways in Clinical Neuropsychiatry 2016:547-571Tosto G, Reitz C. Genomics of Alzheimer's disease: Value of highthroughput genomic technologies to dissect its etiology. Molecular and Cellular Probes 2016;30(6):397-403.Mohamed T, Shakeri A, Rao PPN. Amyloid cascade in Alzheimer's disease: Recent advances in medicinal chemistry. European Journal of Medicinal Chemistry 2016;113:258-272.Sterner RM, Takahashi PY, Yu Ballard AC. Active Vaccines for Alzheimer Disease Treatment. Journal of the American Medical Directors Association 2016;17(9):862.e1-862.e15.López Casillas F, López Casillas F. CRISPR, el sueño divino hecho realidad. Revista de la Facultad de Medicina (México) 2016 08/;58(4):55-60.Kanchiswamy CN, Maffei M, Malnoy M, Velasco R, Kim J. Fine-Tuning Next-Generation Genome Editing Tools. Trends in Biotechnology 2016;34(7):562-574.Maeder ML, Gersbach CA. Genome-editing Technologies for Gene and Cell Therapy. Molecular Therapy 2016;24(3):430-446.Prabhu V, Xu H. Endonuclease mediated genome editing in drug discovery and development: promises and challenges. Drug Discovery Today: Technologies 2016;21-22:17-25.Lander E. The Heroes of CRISPR. Cell 2016;164(1):18-28.Mojica FJM, Montoliu L. On the Origin of CRISPR-Cas Technology: From Prokaryotes to Mammals. Trends in Microbiology 2016;24(10):811-820.Golubov A. Chapter 6 - CRISPR: Bacteria Immune System. Genome Stability 2016:87-98.Han W, She Q. Chapter One - CRISPR History: Discovery, Characterization, and Prosperity. Progress in Molecular Biology and Translational Science 2017;152:1-21Ede DR, Farhang N, Stover JD, Bowles RD. 4.32 Gene Editing Tools. Comprehensive Biomaterials II 2017:589-599.Pelletier S. CRISPR–Cas9. Encyclopedia of Microbiology (Fourth Edition) 2017:772-782.Lemay M, Horvath P, Moineau S. The CRISPR-Cas app goes viral. Current Opinion in Microbiology 2017;37:103-109.Hille F, Richter H, Wong SP, Bratovič M, Ressel S, Charpentier E. The Biology of CRISPR-Cas: Backward and Forward. Cell 2018;172(6):1239-1259Singh V, Braddick D, Dhar PK. Exploring the potential of genome editing CRISPR-Cas9 technology. Gene 2017;599:1-18.Ramirez JC. Chapter Six - Gene Editing and CRISPR Therapeutics: Strategies Taught by Cell and Gene Therapy. Progress in Molecular Biology and Translational Science 2017;152:115-130.Martinez-Lage M, Torres-Ruiz R, Rodriguez-Perales S. Chapter Two - CRISPR/Cas9 Technology: Applications and Human Disease Modeling. Progress in Molecular Biology and Translational Science 2017;152:23-48.Powell SK, Gregory J, Akbarian S, Brennand KJ. Application of CRISPR/Cas9 to the study of brain development and neuropsychiatric disease. Molecular and Cellular Neuroscience 2017;82:157-166.Shrock E, Güell M. Chapter Six - CRISPR in Animals and Animal Models. Progress in Molecular Biology and Translational Science 2017;152:95-114.de Lecuona I, Casado M, Marfany G, Lopez Baroni M, Escarrabill M. Gene Editing in Humans: Towards a Global and Inclusive Debate for Responsible Research. Yale J Biol Med 2017 12;90(4):673-681.Cribbs AP, Perera SMW. Science and Bioethics of CRISPR-Cas9 Gene Editing: An Analysis Towards Separating Facts and Fiction. Yale J Biol Med 2017 12;90(4):625-634.Griffin EF, Caldwell KA, Caldwell GA. Genetic and Pharmacological Discovery for Alzheimer's Disease Using Caenorhabditis elegans. ACS Chem Neurosci 2017 12 20,;8(12):2596-2606.Pir GJ, Choudhary B, Mandelkow E. Caenorhabditis elegans models of tauopathy. FASEB J 2017 12;31(12):5137-5148Ortiz-Virumbrales M, Moreno CL, Kruglikov I, Marazuela P, Sproul A, Jacob S, et al. CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer's PSEN2 N141I neurons. Acta Neuropathol Commun 2017 10 27,;5(1):77.Shao W, Peng D, Wang X. Genetics of Alzheimer’s disease: From pathogenesis to clinical usage. Journal of Clinical Neuroscience 2017;45:1-8.Carmona S, Hardy J, Guerreiro R. Chapter 26 - The genetic landscape of Alzheimer disease. Handbook of Clinical Neurology 2018;148:395-408.Hunter S, Smailagic N, Brayne C. Aβ and the dementia syndrome: Simple versus complex perspectives. European Journal of Clinical Investigation 2018;48(12):e13025.Kim JH. Genetics of Alzheimer's Disease. Dement Neurocogn Disord 2018;17(4):131-136.McDiarmid TA, Au V, Loewen AD, Liang J, Mizumoto K, Moerman DG, et al. CRISPR-Cas9 human gene replacement and phenomic characterization in Caenorhabditis elegans to understand the functional conservation of human genes and decipher variants of uncertain significance. Dis Model Mech 2018 11 26,;11(12).Rohn TT, Kim N, Isho NF, Mack JM. The Potential of CRISPR/Cas9 Gene Editing as a Treatment Strategy for Alzheimer’s Disease. J Alzheimers Dis Parkinsonism 2018;8(3)Rahman S, Datta M, Kim J, Jan AT. CRISPR/Cas: An intriguing genomic editing tool with prospects in treating neurodegenerative diseases. Seminars in Cell & Developmental Biology 2019;96:22-31.Roshni Mishra, Leya Joykutty, Kepa Oyarbide American Heritage School, Plantation, FL. Expression of anti-neurodegeneration genes in mutant Caenorhabditis elegans using CRISPR-Cas9 improves behavior associated with Alzheimer’s Disease 2019.Ins. Europeo de Bioinformática, Ins. Wellcome Trust Sanger, Ins. Ontario para la Investigación del Cáncer. WormBase. Available at: https://wormbase.org, 2020.Caenorhabditis ElegansAlzheimerAβTauCrispr/Cas9LICENSElicense.txtlicense.txttext/plain; charset=utf-814828https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/4/license.txt2f9959eaf5b71fae44bbf9ec84150c7aMD54open accessTEXTCartas derechos de autor.pdf.txtCartas derechos de autor.pdf.txtExtracted texttext/plain24https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/11/Cartas%20derechos%20de%20autor.pdf.txt269ba7e629f8fadbe92d99e25fe0bd8aMD511metadata only accessINFORME FINAL 2020.pdf.txtINFORME FINAL 2020.pdf.txtExtracted texttext/plain134851https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/13/INFORME%20FINAL%202020.pdf.txt8cc25273cd05f20f9985f9e8bf21442dMD513open accessTHUMBNAILCartas derechos de autor.pdf.jpgCartas derechos de autor.pdf.jpgGenerated Thumbnailimage/jpeg9024https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/12/Cartas%20derechos%20de%20autor.pdf.jpg2f830c4815c93690e5d7f0f1564fa2f3MD512metadata only accessINFORME FINAL 2020.pdf.jpgINFORME FINAL 2020.pdf.jpgGenerated Thumbnailimage/jpeg7314https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/14/INFORME%20FINAL%202020.pdf.jpgbb2e472d25e8f261f9caa11497735dd3MD514open accessORIGINALINFORME FINAL 2020.pdfINFORME FINAL 2020.pdfapplication/pdf1680258https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/15/INFORME%20FINAL%202020.pdf36bb7472d24369b848ccfd6fab4d068eMD515open accessCartas derechos de autor.pdfCartas derechos de autor.pdfapplication/pdf104636https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/68/16/Cartas%20derechos%20de%20autor.pdfbfeb2b90137f3c02a4ae30abaf872b85MD516metadata only accessunicolmayor/68oai:repositorio.unicolmayor.edu.co:unicolmayor/682024-04-29 15:26:35.807An error occurred on the license name.|||https://creativecommons.org/licenses/by-nc-sa/4.0/open accessBiblioteca Digital Unicolmayorrepositorio@unicolmayor.edu.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 |