Immunologic Research

, Volume 47, Issue 1–3, pp 172–178 | Cite as

Transcriptional target-based expression cloning of immunoregulatory molecules

  • Rebecca L. Lamason
  • Stefanie M. Lew
  • Joel L. Pomerantz


Immunologic research has benefited tremendously from expression-cloning strategies designed to isolate genes responsible for a wide variety of immunomodulatory activities, including cytokines, receptors, signaling proteins, and transcription factors. Here, we discuss the use of expression-cloning strategies that have been modified to detect cDNAs that influence gene expression as assayed by a transcriptional reporter. We summarize our experience with these screens, review important parameters, and discuss potential modifications.


Expression-cloning Gene discovery NF-κB HIV-1 cDNA library 


  1. 1.
    Pomerantz JL, Denny EM, Baltimore D. CARD11 mediates factor-specific activation of NF-kappaB by the T cell receptor complex. EMBO J. 2002;21:5184–94.CrossRefPubMedGoogle Scholar
  2. 2.
    Hayden MS, West AP, Ghosh S. NF-kappaB and the immune response. Oncogene. 2006;25:6758–80.CrossRefPubMedGoogle Scholar
  3. 3.
    Gaide O, Favier B, Legler DF, Bonnet D, Brissoni B, Valitutti S, et al. CARMA1 is a critical lipid raft-associated regulator of TCR-induced NF-kappa B activation. Nat Immunol. 2002;3:836–43.CrossRefPubMedGoogle Scholar
  4. 4.
    Wang D, You Y, Case SM, McAllister-Lucas LM, Wang L, DiStefano PS, et al. A requirement for CARMA1 in TCR-induced NF-kappa B activation. Nat Immunol. 2002;3:830–5.CrossRefPubMedGoogle Scholar
  5. 5.
    Hara H, Wada T, Bakal C, Kozieradzki I, Suzuki S, Suzuki N, et al. The MAGUK family protein CARD11 is essential for lymphocyte activation. Immunity. 2003;18:763–75.CrossRefPubMedGoogle Scholar
  6. 6.
    Jun JE, Wilson LE, Vinuesa CG, Lesage S, Blery M, Miosge LA, et al. Identifying the MAGUK protein Carma-1 as a central regulator of humoral immune responses and atopy by genome-wide mouse mutagenesis. Immunity. 2003;18:751–62.CrossRefPubMedGoogle Scholar
  7. 7.
    Egawa T, Albrecht B, Favier B, Sunshine MJ, Mirchandani K, O’Brien W, et al. Requirement for CARMA1 in antigen receptor-induced NF-kappa B activation and lymphocyte proliferation. Curr Biol. 2003;13:1252–8.CrossRefPubMedGoogle Scholar
  8. 8.
    Newton K, Dixit VM. Mice lacking the CARD of CARMA1 exhibit defective B lymphocyte development and impaired proliferation of their B and T lymphocytes. Curr Biol. 2003;13:1247–51.CrossRefPubMedGoogle Scholar
  9. 9.
    McCully RR, Pomerantz JL. The protein kinase C-responsive inhibitory domain of CARD11 functions in NF-kappaB activation to regulate the association of multiple signaling cofactors that differentially depend on Bcl10 and MALT1 for association. Mol Cell Biol. 2008;28:5668–86.CrossRefPubMedGoogle Scholar
  10. 10.
    Yang HC, Shen L, Siliciano RF, Pomerantz JL. Isolation of a cellular factor that can reactivate latent HIV-1 without T cell activation. Proc Natl Acad Sci USA. 2009;106:6321–6.CrossRefPubMedGoogle Scholar
  11. 11.
    Palella FJ Jr, Delaney KM, Moorman AC, Loveless MO, Fuhrer J, Satten GA, et al. Declining morbidity and mortality among patients with advanced human immunodeficiency virus infection. HIV Outpatient Study Investigators. N Engl J Med. 1998;338:853–60.CrossRefPubMedGoogle Scholar
  12. 12.
    Siliciano JD, Kajdas J, Finzi D, Quinn TC, Chadwick K, Margolick JB, et al. Long-term follow-up studies confirm the stability of the latent reservoir for HIV-1 in resting CD4+ T cells. Nat Med. 2003;9:727–8.CrossRefPubMedGoogle Scholar
  13. 13.
    Klickstein LB, Neve RL, Golemis EA, Gyuris J. Conversion of mRNA into double-stranded cDNA. Curr Protoc Mol Biol 2008; Chapter 5: Unit5.5.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Rebecca L. Lamason
    • 1
  • Stefanie M. Lew
    • 1
  • Joel L. Pomerantz
    • 1
  1. 1.Department of Biological Chemistry, Institute for Cell Engineering, and Graduate Program in ImmunologyThe Johns Hopkins University School of MedicineBaltimoreUSA

Personalised recommendations