Exploring the Potential of CRISPRa and CRISPRi to Systematically Dissect Fate Options in Hematopoiesis
(2017) KBK820 20171Pure and Applied Biochemistry
- Abstract
- Advances in our understanding of hematopoietic stem cells (HSCs) have enabled the establishment of differentiation pathways throughout the hematopoietic hierarchy. To further explore the molecular cues governing cell fate options, there is a need to manipulate gene function in a controllable manner. Gene manipulation systems based on CRISPR activation (CRISPRa) or interference (CRISPRi) could be used to achieve this requirement. Here, we have generated doxycycline-inducible CRISPRa and CRISPRi mouse models. To functionally validate these systems, we aimed to induce or repress the chosen genes C/EBPβ and Notch1, respectively. An initial requirement to this end was to obtain functional high titer lentiviruses that could be used to deliver... (More)
- Advances in our understanding of hematopoietic stem cells (HSCs) have enabled the establishment of differentiation pathways throughout the hematopoietic hierarchy. To further explore the molecular cues governing cell fate options, there is a need to manipulate gene function in a controllable manner. Gene manipulation systems based on CRISPR activation (CRISPRa) or interference (CRISPRi) could be used to achieve this requirement. Here, we have generated doxycycline-inducible CRISPRa and CRISPRi mouse models. To functionally validate these systems, we aimed to induce or repress the chosen genes C/EBPβ and Notch1, respectively. An initial requirement to this end was to obtain functional high titer lentiviruses that could be used to deliver guide RNAs (gRNAs) to cells from the mouse lines, and work was conducted to optimize the production of lentiviruses. Furthermore, we demonstrate the ability to induce the CRISPRa system. However, initial experiments show difficulties in C/EBPβ mediated transdifferentiation of B cells from the CRISPRa mouse model. Thus, further detailed validations are necessary to address the potential problems. (Less)
- Popular Abstract
- A bacterial immune system called CRISPR can be modified to overexpress or repress any gene of interest, which makes it possible to study why some blood cells develop normally while others become diseased.
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/8913760
- author
- Bäckström, Alexandra LU
- supervisor
-
- David Bryder LU
- Amol Ugale LU
- Eva Erlandsson LU
- organization
- course
- KBK820 20171
- year
- 2017
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- Applied biochemistry, tillämpad biokemi
- language
- English
- id
- 8913760
- date added to LUP
- 2017-09-14 15:41:17
- date last changed
- 2017-09-14 15:41:17
@misc{8913760, abstract = {{Advances in our understanding of hematopoietic stem cells (HSCs) have enabled the establishment of differentiation pathways throughout the hematopoietic hierarchy. To further explore the molecular cues governing cell fate options, there is a need to manipulate gene function in a controllable manner. Gene manipulation systems based on CRISPR activation (CRISPRa) or interference (CRISPRi) could be used to achieve this requirement. Here, we have generated doxycycline-inducible CRISPRa and CRISPRi mouse models. To functionally validate these systems, we aimed to induce or repress the chosen genes C/EBPβ and Notch1, respectively. An initial requirement to this end was to obtain functional high titer lentiviruses that could be used to deliver guide RNAs (gRNAs) to cells from the mouse lines, and work was conducted to optimize the production of lentiviruses. Furthermore, we demonstrate the ability to induce the CRISPRa system. However, initial experiments show difficulties in C/EBPβ mediated transdifferentiation of B cells from the CRISPRa mouse model. Thus, further detailed validations are necessary to address the potential problems.}}, author = {{Bäckström, Alexandra}}, language = {{eng}}, note = {{Student Paper}}, title = {{Exploring the Potential of CRISPRa and CRISPRi to Systematically Dissect Fate Options in Hematopoiesis}}, year = {{2017}}, }