A lentiviral screen for novel regulators of IL-7R[alpha] in a pre-B cell line

IL-7R[alpha] is one component of the heterodimeric IL-7R[alpha] receptor, and signaling through this receptor is essential for murine T and B cell development as well as human T cell development. IL-7R[alpha] signaling is also responsible for homeostatic proliferation of T cells in lymphopenic hosts, as well as maintenance of naïve and memory CD8+ T cells in the periphery. A number of regulators of IL-7R[alpha][alpha] have been identified, but the complex processes underlying fine control of IL-7R[alpha][alpha] expression are poorly understood. The RNAi Consortium's lentivirus-based shRNA library targeting murine kinases and phosphatases has allowed large scale screening for modulators of IL-7R[alpha][alpha] surface expression. This library provides shRNAs in the pLKO. 1 lentiviral vector targeting 1278 known kinases and phosphatases. Analysis of the FACS-based assay identified 38 potential regulators of IL-7R[alpha][alpha] in a pre-B cell line. Subsequent validation of five of the hits confirmed known pathways of IL-7R regulation and also pointed to new potential points of regulatory control. Phosphoinositide 3-kinase (PI3K) regulates a number of cell signaling pathways that promote survival, proliferation and increased metabolism. In mammals the Class I family of PI3Ks consists of four different catalytic subunits, which can pair with any of six different regulatory subunits. PI3K was recently shown to regulate the expression of the interleukin-7 receptor alpha chain (IL-7R[alpha][alpha]) via the Foxol transcription factor.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Biology.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2009

Author dc:creator

• Trajman, Lily Christine

Advisor dc:contributor.advisor

• Jianzhu Chen.

• Biology.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/47885

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/47885