miR-290 cluster (red) and miR-302 cluster (green) expression during mouse embryonic development. Cells within dotted line are embryonic tissue and outside of dotted line are extraembryonic tissue. Sorting experiments confirmed that the yellow represents expression of both clusters in the same cell.

Two miRNA Clusters Reveal Alternative Paths in Late-Stage Reprogramming

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Figure 1b in this paper is one my favorite images. Prior to this study, we had demonstrated that when overexpressed, microRNAs from the miR-290 and miR-302 clusters enhance the efficiency and specificity of reprogramming somatic cells to a pluripotent state, and other labs had demonstrated that the miR-302 cluster can even induce iPSC formation on its own. Motivated by this powerful phenotype, Ron Parchem wanted to investigate the role of the endogenous clusters during both reprogramming and normal development. He constructed a beautiful system by generating live fluorescent reporters for each cluster. Figure 1b shows the expression of these reporters during mouse embryonic development, highlighting his fascinating discovery that the expression of the two clusters can be used to mark not only the naive and primed stem cells of the blastocyst and epiblast, respectively, but also a transient third pluripotent state where both clusters are active. This ordered differentiation also occurs during in vitro stem cell differentiation. In contrast, although the endogenous activation of either cluster is predictive of successful downstream reprogramming, the process of dedifferentiation is not similarly ordered, but can take alternative paths.

Model of three states of pluripotency, a naive state (red), a primed state (green), and an intermediate state (yellow). Differentiation both during development and in vivo passes through each state in an ordered manner. During de-differentiation, however, there are alternatvie routes to reach the the naive (red) state, that do not required passing back through the primed or intermediate pluripotency states.

Publications

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Combined activation of MAP kinase pathway and β-catenin signaling cause deep penetrating nevi.

Nature Communications accepted 2017

Iwei Yeh, Ursula Lang, Emeline Durieux, Meng Kian Tee, Aparna Jorapur, Alan Shain, Veronique Haddad, Daniel Pissaloux, Xu Chen, Lorenzo C...

High accuracy label-free classification of single cell kinetic states from holographic cytometry of human melanoma cells.

Scientific Reports accepted 2017

Miro Hejna, Aparna Jorpaur, Jun Song, Robert L Judson.

CDK1 Inhibition Targets the p53-NOXA-MCL1 Axis, Selectively Kills Embryonic Stem Cells, and Prevents Teratoma Formation

Stem Cell Reports 2015

Noelle E Huskey, Tingxia Guo, Kimberley J Evason, Olga Momcilovic, David Pardo, Katelyn J Creasman, Robert L Judson, Robert Blelloch, Sco...

Two miRNA Clusters Reveal Alternative Paths in Late-Stage Reprogramming

Cell Stem Cell 2014

Ronald J Parchem, Julia Ye, Robert L Judson, Marie F Larussa,Raga Krishnakumar, Amy Blelloch, Michael C Oldham, Robert Blelloch

MicroRNA-based discovery of barriers to dedifferentiation of fibroblasts to pluripotent stem cells

Nature Structural & Molecular Biology 2013

Robert L Judson, Tobias S Greve, Ronald J Parchem, Robert Blelloch

microRNA Control of Mouse and Human Pluripotent Stem Cell Behavior

Annual Review of Cell and Developmental Biology 2013

Tobias S Greve, Robert L Judson, Robert Blelloch

Multiple targets of miR-302 and miR-372 promote reprogramming of human fibroblasts to induced pluripotent stem cells

Nature Biotechnology 2011

Deepa Subramanyam, Samy Lamouille, Robert L Judson, Jason Y Liu, Nathan Bucay, Rik Derynck, Robert Blelloch

miR-380-5p represses p53 to control cellular survival and is associated with poor outcome in MYCN-amplified neuroblastoma

Nature Medicine 2010

Alexander Swarbrick, Susan L Woods, Alexander Shaw, Asha Balakrishnan, Yuwei Phua, Akira Nguyen, Yvan Chanthery, Lionel Lim, Lesley J Ash...

Opposing microRNA families regulate self-renewal in mouse embryonic stem cells

Nature 2010

Collin Melton, Robert L Judson, Robert Blelloch

Embryonic stem cell-specific microRNAs promote induced pluripotency

Nature Biotechnology 2009

Robert L Judson, Joshua E Babiarz, Monica Venere, Robert Blelloch

The GP(Y/F) domain of TF1 integrase multimerizes when present in a fragment, and substitutions in this domain reduce enzymatic activity of the full-length protein

Journal of Biological Chemistry 2008

Hirotaka Ebina, Atreyi Ghatak Chatterjee, Robert L Judson, Henry L Levin

The self primer of the long terminal repeat retrotransposon Tf1 is not removed during reverse transcription.

Journal of Virology 2006

Angela Atwood-Moore, Kenneth Yan, Robert L Judson, Henry L Levin

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