An emerging technology known as three-dimensional (3D) tissue engineering has allowed scientists to mimic tissues found in vivo. Previous studies indicate that it is possible to differentiate dissociated mouse embryonic stem cells (mESCs) into 3D retinal tissues in vitro (Bertacchi, 2015; Eiraku, 2012). The newly differentiated retinal tissues are said to encompass all of the major components found in retinal tissues. The generation of in vitro 3D tissues holds great potential in terms of patient-specific disease modeling. Although various diseases have been well-studied in animal models, there are limitations with regards to patient-specificity. The generation of animal models to study each specific mutation would be impractical and costly. With in vitro generation of tissues and organs, scientists have the capacity to generate disease models consisting of mutations specific to patients.
There are various gene editing techniques available to generate and recreate disease-specific 3D disease models using stem cells. The newest gene editing technique known as Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR) has allowed scientists to edit the genome with greater precision than other genome editing technologies like Zinc Finger Nucleases (ZFNs) or Transcription Activator-like Effector Nucleases (TALENs).
In order to overcome current limitations on disease-specific model systems, we developed a protocol for differentiating mouse embryonic stem cells into 3D retinas in vitro. In addition, to confirm the utility of this 3D model for future disease-specific mutations, we utilized the CRISPR method as a means to knockdown the transcription factor CRX, which is known to regulate key developmental decisions during retinogenesis, and as such, mutations in CRX have resulted in various diseases such as Leber Congenital Amaurosis, Cone-Rod Dystrophy, and Retinitis Pigmentosa.
","abstract_html":"<p>An emerging technology known as three-dimensional (3D) tissue engineering has allowed scientists to mimic tissues found <em>in vivo.</em> Previous studies indicate that it is possible to differentiate dissociated mouse embryonic stem cells (mESCs) into 3D retinal tissues <em>in vitro</em><em> </em>(Bertacchi, 2015; Eiraku, 2012). The newly differentiated retinal tissues are said to encompass all of the major components found in retinal tissues. The generation of <em>in vitro</em> 3D tissues holds great potential in terms of patient-specific disease modeling. Although various diseases have been well-studied in animal models, there are limitations with regards to patient-specificity. The generation of animal models to study each specific mutation would be impractical and costly. With <em>in vitro</em> generation of tissues and organs, scientists have the capacity to generate disease models consisting of mutations specific to patients.</p> <p>There are various gene editing techniques available to generate and recreate disease-specific 3D disease models using stem cells. The newest gene editing technique known as Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR) has allowed scientists to edit the genome with greater precision than other genome editing technologies like Zinc Finger Nucleases (ZFNs) or Transcription Activator-like Effector Nucleases (TALENs).</p> <p>In order to overcome current limitations on disease-specific model systems, we developed a protocol for differentiating mouse embryonic stem cells into 3D retinas <em>in vitro</em>. In addition, to confirm the utility of this 3D model for future disease-specific mutations, we utilized the CRISPR method as a means to knockdown the transcription factor CRX, which is known to regulate key developmental decisions during retinogenesis, and as such, mutations in CRX have resulted in various diseases such as Leber Congenital Amaurosis, Cone-Rod Dystrophy, and Retinitis Pigmentosa.</p>","abstract_has_math":false,"creators":["Prasad, Pooja"],"institution":null,"degree_name":"Master of Science","degree_level":"Master's Thesis","degree_discipline":"Biological Sciences","degree_department":null,"school":null,"contributors":["Deepak Lamba, MBBS, PhD","Mary B. Sevigny, PhD"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2017,"date_issued":"2017-05-01T07:00:00Z","date_published":"2017-05-01T07:00:00Z","updated_at":"2026-07-24T02:04:44Z","subjects":["CRISPR","Retina","Organoid","Crx","Mouse Embryonic Stem Cells","Knockdown","Biology","Developmental Biology","Developmental Neuroscience","Molecular and Cellular Neuroscience","Molecular Genetics"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://scholar.dominican.edu/masters-theses/279","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Deepak Lamba, MBBS, PhD","Mary B. Sevigny, PhD"]},{"key":"dc:creator","label":"Author","values":["Prasad, Pooja"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2020-06-01T07:00:00Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biological Sciences"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Master's Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Master of Science"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["CRISPR","Retina","Organoid","Crx","Mouse Embryonic Stem Cells","Knockdown","Biology","Developmental Biology","Developmental Neuroscience","Molecular and Cellular Neuroscience","Molecular Genetics"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://scholar.dominican.edu/masters-theses/279"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["An emerging technology known as three-dimensional (3D) tissue engineering has allowed scientists to mimic tissues found in vivo. Previous studies indicate that it is possible to differentiate dissociated mouse embryonic stem cells (mESCs) into 3D retinal tissues in vitro (Bertacchi, 2015; Eiraku, 2012). The newly differentiated retinal tissues are said to encompass all of the major components found in retinal tissues. The generation of in vitro 3D tissues holds great potential in terms of patient-specific disease modeling. Although various diseases have been well-studied in animal models, there are limitations with regards to patient-specificity. The generation of animal models to study each specific mutation would be impractical and costly. With in vitro generation of tissues and organs, scientists have the capacity to generate disease models consisting of mutations specific to patients.
There are various gene editing techniques available to generate and recreate disease-specific 3D disease models using stem cells. The newest gene editing technique known as Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR) has allowed scientists to edit the genome with greater precision than other genome editing technologies like Zinc Finger Nucleases (ZFNs) or Transcription Activator-like Effector Nucleases (TALENs).
In order to overcome current limitations on disease-specific model systems, we developed a protocol for differentiating mouse embryonic stem cells into 3D retinas in vitro. In addition, to confirm the utility of this 3D model for future disease-specific mutations, we utilized the CRISPR method as a means to knockdown the transcription factor CRX, which is known to regulate key developmental decisions during retinogenesis, and as such, mutations in CRX have resulted in various diseases such as Leber Congenital Amaurosis, Cone-Rod Dystrophy, and Retinitis Pigmentosa.
"]},{"key":"dc:title","label":"Title","values":["Modeling 3D Retinogenesis in Mouse Embryonic Stem Cells Following CRISPR-mediated Crx Knockdown"]}]}],"canonical_facts":{"dc:contributor":["Deepak Lamba, MBBS, PhD","Mary B. Sevigny, PhD"],"dc:creator":["Prasad, Pooja"],"dc:date.available":["2020-06-01T07:00:00Z"],"dc:description.abstract":["An emerging technology known as three-dimensional (3D) tissue engineering has allowed scientists to mimic tissues found in vivo. Previous studies indicate that it is possible to differentiate dissociated mouse embryonic stem cells (mESCs) into 3D retinal tissues in vitro (Bertacchi, 2015; Eiraku, 2012). The newly differentiated retinal tissues are said to encompass all of the major components found in retinal tissues. The generation of in vitro 3D tissues holds great potential in terms of patient-specific disease modeling. Although various diseases have been well-studied in animal models, there are limitations with regards to patient-specificity. The generation of animal models to study each specific mutation would be impractical and costly. With in vitro generation of tissues and organs, scientists have the capacity to generate disease models consisting of mutations specific to patients.
There are various gene editing techniques available to generate and recreate disease-specific 3D disease models using stem cells. The newest gene editing technique known as Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR) has allowed scientists to edit the genome with greater precision than other genome editing technologies like Zinc Finger Nucleases (ZFNs) or Transcription Activator-like Effector Nucleases (TALENs).
In order to overcome current limitations on disease-specific model systems, we developed a protocol for differentiating mouse embryonic stem cells into 3D retinas in vitro. In addition, to confirm the utility of this 3D model for future disease-specific mutations, we utilized the CRISPR method as a means to knockdown the transcription factor CRX, which is known to regulate key developmental decisions during retinogenesis, and as such, mutations in CRX have resulted in various diseases such as Leber Congenital Amaurosis, Cone-Rod Dystrophy, and Retinitis Pigmentosa.
"],"dc:identifier":["https://scholar.dominican.edu/masters-theses/279"],"dc:subject":["CRISPR","Retina","Organoid","Crx","Mouse Embryonic Stem Cells","Knockdown","Biology","Developmental Biology","Developmental Neuroscience","Molecular and Cellular Neuroscience","Molecular Genetics"],"dc:title":["Modeling 3D Retinogenesis in Mouse Embryonic Stem Cells Following CRISPR-mediated Crx Knockdown"],"thesis:degree_discipline":["Biological Sciences"],"thesis:degree_level":["Master's Thesis"],"thesis:degree_name":["Master of Science"]},"updated_at":"2026-07-24T02:04:44Z"}