Cas9 effector-mediated regulation of transcription and differentiation in human pluripotent stem cells
Diabetes Center of Excellence; Program in Molecular Medicine; Program in Gene Function and Expression; Bioinformatics and Integrative Biology
Medical Subject Headings
Amino Acid Sequence; Caspase 9; Cell Differentiation; Cell Lineage; Clustered Regularly Interspaced Short Palindromic Repeats; DNA-Binding Proteins; Embryonic Stem Cells; Gene Regulatory Networks; HEK293 Cells; Humans; Molecular Sequence Data; Octamer Transcription Factor-3; Pluripotent Stem Cells; SOXF Transcription Factors; Transcription, Genetic; Transcriptional Activation
Cell Biology | Cellular and Molecular Physiology | Developmental Biology | Molecular Genetics
The identification of the trans-acting factors and cis-regulatory modules that are involved in human pluripotent stem cell (hPSC) maintenance and differentiation is necessary to dissect the operating regulatory networks in these processes and thereby identify nodes where signal input will direct desired cell fate decisions in vitro or in vivo. To deconvolute these networks, we established a method to influence the differentiation state of hPSCs with a CRISPR-associated catalytically inactive dCas9 fused to an effector domain. In human embryonic stem cells, we find that the dCas9 effectors can exert positive or negative regulation on the expression of developmentally relevant genes, which can influence cell differentiation status when impinging on a key node in the regulatory network that governs the cell state. This system provides a platform for the interrogation of the underlying regulators governing specific differentiation decisions, which can then be employed to direct cellular differentiation down desired pathways.
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Citation: Kearns NA, Genga RM, Enuameh MS, Garber M, Wolfe SA, Maehr R. Cas9 effector-mediated regulation of transcription and differentiation in human pluripotent stem cells. Development. 2014 Jan;141(1):219-23. doi:10.1242/dev.103341. Link to article on publisher's site
Kearns, Nicola A.; Genga, Ryan M. J.; Enuameh, Metewo Selase; Garber, Manuel; Wolfe, Scot A.; and Maehr, Rene, "Cas9 effector-mediated regulation of transcription and differentiation in human pluripotent stem cells" (2014). University of Massachusetts Medical School Faculty Publications. 337.