An inducible cell model for studies of V(D)J recombinational control

Michael L. Sikes; Eugene M. Oltz

doi:10.1016/S0022-1759(99)00003-4

An inducible cell model for studies of V(D)J recombinational control

Michael L. Sikes, Eugene M. Oltz

Division of Laboratory and Genomic Medicine

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

Antigen receptor gene assembly is controlled by enhancer-directed changes in the accessibility of chromosomal gene segments to V(D)J recombinase. To dissect mechanisms that regulate rearrangement efficiencies, we developed a cell system (TDR19) in which recombination activating gene (RAG) expression is repressed by tetracycline. Under conditions of RAG repression, recombination substrates were consistently integrated into the TDR19 genome in an unrearranged form. Subsequent rearrangement of chromosomal substrates containing a transcriptional enhancer correlated inversely with tetracycline concentrations. Together, these features underscore the utility of TDR19 as a cell model for defining the molecular determinants of V(D)J recombinational accessibility.

Original language	English
Pages (from-to)	25-29
Number of pages	5
Journal	Journal of Immunological Methods
Volume	224
Issue number	1-2
DOIs	https://doi.org/10.1016/S0022-1759(99)00003-4
State	Published - Apr 22 1999

Keywords

Inducible promoter
T Cell receptor
Transcriptional enhancers
V(D)J recombination

Access to Document

10.1016/S0022-1759(99)00003-4

Cite this

@article{8ba10dea5d8048dcae6c3b2cbcc5bcde,

title = "An inducible cell model for studies of V(D)J recombinational control",

abstract = "Antigen receptor gene assembly is controlled by enhancer-directed changes in the accessibility of chromosomal gene segments to V(D)J recombinase. To dissect mechanisms that regulate rearrangement efficiencies, we developed a cell system (TDR19) in which recombination activating gene (RAG) expression is repressed by tetracycline. Under conditions of RAG repression, recombination substrates were consistently integrated into the TDR19 genome in an unrearranged form. Subsequent rearrangement of chromosomal substrates containing a transcriptional enhancer correlated inversely with tetracycline concentrations. Together, these features underscore the utility of TDR19 as a cell model for defining the molecular determinants of V(D)J recombinational accessibility.",

keywords = "Inducible promoter, T Cell receptor, Transcriptional enhancers, V(D)J recombination",

author = "Sikes, {Michael L.} and Oltz, {Eugene M.}",

note = "Funding Information: We thank Dr. David Schatz (Yale Univ.) for the pTET-R1 and pTET-R2 constructs, and for anti-RAG-1 and anti-RAG-2 antibodies. This work was supported by NIH grants AI36944, AI01412 (EMO) and GM19597 (MLS). EMO is a Joe C. Davis Scholar. ",

year = "1999",

month = apr,

day = "22",

doi = "10.1016/S0022-1759(99)00003-4",

language = "English",

volume = "224",

pages = "25--29",

journal = "Journal of Immunological Methods",

issn = "0022-1759",

number = "1-2",

}

TY - JOUR

T1 - An inducible cell model for studies of V(D)J recombinational control

AU - Sikes, Michael L.

AU - Oltz, Eugene M.

N1 - Funding Information: We thank Dr. David Schatz (Yale Univ.) for the pTET-R1 and pTET-R2 constructs, and for anti-RAG-1 and anti-RAG-2 antibodies. This work was supported by NIH grants AI36944, AI01412 (EMO) and GM19597 (MLS). EMO is a Joe C. Davis Scholar.

PY - 1999/4/22

Y1 - 1999/4/22

N2 - Antigen receptor gene assembly is controlled by enhancer-directed changes in the accessibility of chromosomal gene segments to V(D)J recombinase. To dissect mechanisms that regulate rearrangement efficiencies, we developed a cell system (TDR19) in which recombination activating gene (RAG) expression is repressed by tetracycline. Under conditions of RAG repression, recombination substrates were consistently integrated into the TDR19 genome in an unrearranged form. Subsequent rearrangement of chromosomal substrates containing a transcriptional enhancer correlated inversely with tetracycline concentrations. Together, these features underscore the utility of TDR19 as a cell model for defining the molecular determinants of V(D)J recombinational accessibility.

AB - Antigen receptor gene assembly is controlled by enhancer-directed changes in the accessibility of chromosomal gene segments to V(D)J recombinase. To dissect mechanisms that regulate rearrangement efficiencies, we developed a cell system (TDR19) in which recombination activating gene (RAG) expression is repressed by tetracycline. Under conditions of RAG repression, recombination substrates were consistently integrated into the TDR19 genome in an unrearranged form. Subsequent rearrangement of chromosomal substrates containing a transcriptional enhancer correlated inversely with tetracycline concentrations. Together, these features underscore the utility of TDR19 as a cell model for defining the molecular determinants of V(D)J recombinational accessibility.

KW - Inducible promoter

KW - T Cell receptor

KW - Transcriptional enhancers

KW - V(D)J recombination

UR - http://www.scopus.com/inward/record.url?scp=0033025470&partnerID=8YFLogxK

U2 - 10.1016/S0022-1759(99)00003-4

DO - 10.1016/S0022-1759(99)00003-4

M3 - Article

C2 - 10357203

AN - SCOPUS:0033025470

SN - 0022-1759

VL - 224

SP - 25

EP - 29

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

IS - 1-2

ER -

An inducible cell model for studies of V(D)J recombinational control

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this