Reference - Detail
RRC ID | 1522 |
---|---|
Author | Kanayama N, Todo K, Reth M, Ohmori H. |
Title | Reversible switching of immunoglobulin hypermutation machinery in a chicken B cell line. |
Journal | Biochem Biophys Res Commun |
Abstract |
A chicken B lymphoma line, DT40, hypermutates immunoglobulin (Ig) genes spontaneously during culture. Thus, cultured DT 40 cells constitute a useful Ig library for screening antibodies (Abs) in vitro. To fix desirable Ig mutants by stopping hypermutation or to resume mutation for further improvement of Ab affinity, activation-induced cytidine deaminase (AID), a key enzyme responsible for the Ig mutation machinery, must be switched on or off. To this end, we generated a DT40 line whose one AID allele was disrupted, and the other allele was replaced by the loxP-flanked AID construct. In this engineered cell line designated as DT40-SW, AID expression could be switched reversibly by tamoxifen-regulated Cre recombinase. Devices were also introduced to discriminate between the "AID-ON" and the "AID-OFF" cells by GFP expression and puromycin resistance, respectively. Starting from a single DT40-SW cell, Ig gene repertoire was efficiently diversified during culture only when AID expression was on. |
Volume | 327(1) |
Pages | 70-5 |
Published | 2005-2-4 |
DOI | 10.1016/j.bbrc.2004.11.143 |
PII | S0006-291X(04)02753-6 |
PMID | 15629431 |
MeSH | Animals Attachment Sites, Microbiological / genetics B-Lymphocytes / metabolism* B-Lymphocytes / pathology Cell Line, Tumor Chickens / genetics* Chickens / immunology Cytidine Deaminase / genetics* Cytidine Deaminase / metabolism* Drug Resistance / genetics Green Fluorescent Proteins / genetics Hydroxytestosterones / pharmacology Immunoglobulins / genetics* Puromycin / pharmacology Somatic Hypermutation, Immunoglobulin / genetics* Tamoxifen / analogs & derivatives* Tamoxifen / pharmacology |
IF | 2.985 |
Times Cited | 13 |
WOS Category | BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY |
Resource | |
Human and Animal Cells | DT40(RCB1464) |