FAMILY REHAB

FAMILY REHAB

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Large-scale in vivo CRISPR screens identify SAGA complex members as a key regulators of HSC lineage commitment and aging | bioRxiv
Large-scale in vivo CRISPR screens identify SAGA complex members as a key regulators of HSC lineage commitment and aging | bioRxiv
The biological mechanisms that sustain the vast blood production required for healthy life remain incompletely understood. To address this knowledge gap, we developed an in vivo hematopoietic stem cell (HSC)-based large-scale CRISPR knockout screening platform to enable the genetic interrogation of hematopoiesis and broad aspects of immune cell function in vivo. Targeting ∼7000 genes with this methodology, we discovered SAGA complex members Tada2b and Taf5l as key regulators of HSC lineage commitment. Loss of Tada2b or Taf5l inhibited hematopoiesis in vivo and was associated with upregulation of interferon response gene expression. SAGA complex member expression is significantly reduced in aged HSCs and upregulated with heterochronic parabiosis, suggesting a novel mechanism of age-associated hematopoietic decline and rejuvenation. Our study provides a rich functional genetics resource of hematopoiesis regulators accessible through a public interactive database ([www.hematopoiesiscrisprscreens.com][1]), a novel mechanism regulating age-related decline of hematopoiesis, and a new methodology with broad applications to systematically probe the development and functions of the lymphohematopoietic system. ### Competing Interest Statement H.N. is a co-founder and shareholder in ReproCELL, Megakaryon, and Century Therapeutics. [1]: http://www.hematopoiesiscrisprscreens.com
·biorxiv.org·
Large-scale in vivo CRISPR screens identify SAGA complex members as a key regulators of HSC lineage commitment and aging | bioRxiv
Rapid characterization of CRISPR-Cas9 protospacer adjacent motif sequence elements | Genome Biology | Full Text
Rapid characterization of CRISPR-Cas9 protospacer adjacent motif sequence elements | Genome Biology | Full Text
To expand the repertoire of Cas9s available for genome targeting, we present a new in vitro method for the simultaneous examination of guide RNA and protospacer adjacent motif (PAM) requirements. The method relies on the in vitro cleavage of plasmid libraries containing a randomized PAM as a function of Cas9-guide RNA complex concentration. Using this method, we accurately reproduce the canonical PAM preferences for Streptococcus pyogenes, Streptococcus thermophilus CRISPR3 (Sth3), and CRISPR1 (Sth1). Additionally, PAM and sgRNA solutions for a novel Cas9 protein from Brevibacillus laterosporus are provided by the assay and are demonstrated to support functional activity in vitro and in plants.
·genomebiology.biomedcentral.com·
Rapid characterization of CRISPR-Cas9 protospacer adjacent motif sequence elements | Genome Biology | Full Text
Unregulated Contaminant Monitoring Rule | Florida Department of Environmental Protection
Unregulated Contaminant Monitoring Rule | Florida Department of Environmental Protection
What is the Unregulated Contaminant Monitoring Rule?The U.S. Environmental Protection Agency (EPA) periodically requires public water systems throughout the nation to conduct monitoring under the Unregulated Contaminant Monitoring Rule (UCMR) for contaminants that may be present in drinking water but are not currently subjected to federal drinking water regulations.EPA
·floridadep.gov·
Unregulated Contaminant Monitoring Rule | Florida Department of Environmental Protection
Make Home Life Private Again
Make Home Life Private Again
The sacred nature of what we do compels us to privatize our lives, because only then can we be fully invested in making the most of them.
·thefederalist.com·
Make Home Life Private Again
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