Temporal Requirements of cMyc Protein for Reprogramming Mouse Fibroblasts
Exogenous expression of Oct4, Sox2, Klf4, and cMyc forces mammalian somatic cells to adopt molecular and phenotypic characteristics of embryonic stem cells, commencing with the required suppression of lineage-associated genes (e.g., Thy1 in mouse). Although omitting cMyc from the reprogramming cockt...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2012-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2012/541014 |
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| _version_ | 1832568163897704448 |
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| author | Corey Heffernan Huseyin Sumer Luis F. Malaver-Ortega Paul J. Verma |
| author_facet | Corey Heffernan Huseyin Sumer Luis F. Malaver-Ortega Paul J. Verma |
| author_sort | Corey Heffernan |
| collection | DOAJ |
| description | Exogenous expression of Oct4, Sox2, Klf4, and cMyc forces mammalian somatic cells to adopt molecular and phenotypic characteristics of embryonic stem cells, commencing with the required suppression of lineage-associated genes (e.g., Thy1 in mouse). Although omitting cMyc from the reprogramming cocktail minimizes risks of uncontrolled proliferation, its exclusion results in fold reductions in reprogramming efficiency. Thus, the feasibility of substituting cMyc transgene with (non-integrative) recombinant “pTAT-mcMyc” protein delivery was assessed, without compromising reprogramming efficiency or the pluripotent phenotype. Purification and delivery of semisoluble/particulate pTAT-mcMyc maintained Oct4-GFP+ colony formation (i.e., reprogramming efficiency) whilst supporting pluripotency by various criteria. Differential repression of Thy1 by pTAT-mcMyc ± Oct4, Sox2, and Klf4 (OSK) suggested differential (and non-additive) mechanisms of repression. Extending these findings, attempts to enhance reprogramming efficiency through a staggered approach (prerepression of Thy1) failed to improve reprogramming efficiency. We consider protein delivery a useful tool to decipher temporal/molecular events characterizing somatic cell reprogramming. |
| format | Article |
| id | doaj-art-aa954a2f71af46d5ac3d8489831fac39 |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-aa954a2f71af46d5ac3d8489831fac392025-02-03T00:59:40ZengWileyStem Cells International1687-966X1687-96782012-01-01201210.1155/2012/541014541014Temporal Requirements of cMyc Protein for Reprogramming Mouse FibroblastsCorey Heffernan0Huseyin Sumer1Luis F. Malaver-Ortega2Paul J. Verma3Cell Reprogramming and Stem Cells Laboratory, Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, 27-31 Wright Street, Clayton, VIC 3168, AustraliaCell Reprogramming and Stem Cells Laboratory, Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, 27-31 Wright Street, Clayton, VIC 3168, AustraliaCell Reprogramming and Stem Cells Laboratory, Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, 27-31 Wright Street, Clayton, VIC 3168, AustraliaCell Reprogramming and Stem Cells Laboratory, Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, 27-31 Wright Street, Clayton, VIC 3168, AustraliaExogenous expression of Oct4, Sox2, Klf4, and cMyc forces mammalian somatic cells to adopt molecular and phenotypic characteristics of embryonic stem cells, commencing with the required suppression of lineage-associated genes (e.g., Thy1 in mouse). Although omitting cMyc from the reprogramming cocktail minimizes risks of uncontrolled proliferation, its exclusion results in fold reductions in reprogramming efficiency. Thus, the feasibility of substituting cMyc transgene with (non-integrative) recombinant “pTAT-mcMyc” protein delivery was assessed, without compromising reprogramming efficiency or the pluripotent phenotype. Purification and delivery of semisoluble/particulate pTAT-mcMyc maintained Oct4-GFP+ colony formation (i.e., reprogramming efficiency) whilst supporting pluripotency by various criteria. Differential repression of Thy1 by pTAT-mcMyc ± Oct4, Sox2, and Klf4 (OSK) suggested differential (and non-additive) mechanisms of repression. Extending these findings, attempts to enhance reprogramming efficiency through a staggered approach (prerepression of Thy1) failed to improve reprogramming efficiency. We consider protein delivery a useful tool to decipher temporal/molecular events characterizing somatic cell reprogramming.http://dx.doi.org/10.1155/2012/541014 |
| spellingShingle | Corey Heffernan Huseyin Sumer Luis F. Malaver-Ortega Paul J. Verma Temporal Requirements of cMyc Protein for Reprogramming Mouse Fibroblasts Stem Cells International |
| title | Temporal Requirements of cMyc Protein for Reprogramming Mouse Fibroblasts |
| title_full | Temporal Requirements of cMyc Protein for Reprogramming Mouse Fibroblasts |
| title_fullStr | Temporal Requirements of cMyc Protein for Reprogramming Mouse Fibroblasts |
| title_full_unstemmed | Temporal Requirements of cMyc Protein for Reprogramming Mouse Fibroblasts |
| title_short | Temporal Requirements of cMyc Protein for Reprogramming Mouse Fibroblasts |
| title_sort | temporal requirements of cmyc protein for reprogramming mouse fibroblasts |
| url | http://dx.doi.org/10.1155/2012/541014 |
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