Stable gene expression from a mammalian artificial chromosome

被引：73

作者：

Grimes, BR ^{[1
]}

Schindelhauer, D ^{[1
]}

McGill, NI ^{[1
]}

Ross, A ^{[1
]}

Ebersole, TA ^{[1
]}

Cooke, HJ ^{[1
]}

机构：

[1] Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland

来源：

EMBO REPORTS | 2001年 / 2卷 / 10期

关键词：

D O I：

10.1093/embo-reports/kve187

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We have investigated the potential of PAC-based vectors as a route to the incorporation of a gene in a mammalian artificial chromosome (MAC). Previously we demonstrated that a PAC (PAC7c5) containing a-satellite DNA generated mitotically stable MACs in human cells. To determine whether a functional HPRT gene could be assembled in a MAC, PAC7c5 was cotransfected with a second PAC containing a 140 kb human HPRT gene into HPRT-deficient HT1080 cells. Lines were isolated containing a MAC hybridizing with both alpha -satellite and HPRT probes. The MACs segregated efficiently, associated with kinetochore proteins and stably expressed HPRT message after 60 days without selection. Complementation of the parental HPRT deficiency was confirmed phenotypically by growth on HAT selection. These results suggest that MACs could be further developed for delivering a range of genomic copies of genes into cells and that stable transgene expression can be achieved.

引用

页码：910 / 914

页数：5

共 15 条

[1]

BAYNE RAL, 1994, HUM MOL GENET, V3, P539

[2] Mammalian artificial chromosome formation from circular alphoid input DNA does not require telomere repeats [J].

Ebersole, TA ;

Ross, A ;

Clark, E ;

McGill, N ;

Schindelhauer, D ;

Cooke, H ;

Grimes, B .

HUMAN MOLECULAR GENETICS, 2000, 9 (11) :1623-1631

[3] FUNCTIONAL REINTRODUCTION OF HUMAN TELOMERES INTO MAMMALIAN-CELLS [J].

FARR, C ;

FANTES, J ;

GOODFELLOW, P ;

COOKE, H .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (16) :7006-7010

[4] Locus control region function and heterochromatin-induced position effect variegation [J].

Festenstein, R ;

Tolaini, M ;

Corbella, P ;

Mamalaki, C ;

Parrington, J ;

Fox, M ;

Miliou, A ;

Jones, M ;

Kioussis, D .

SCIENCE, 1996, 271 (5252) :1123-1125

[5] IDENTIFICATION OF A SINGLE NUCLEOTIDE CHANGE IN THE HYPOXANTHINE-GUANINE PHOSPHORIBOSYLTRANSFERASE GENE (HPRTYALE) RESPONSIBLE FOR LESCH-NYHAN SYNDROME [J].

FUJIMORI, S ;

DAVIDSON, BL ;

KELLEY, WN ;

PALELLA, TD .

JOURNAL OF CLINICAL INVESTIGATION, 1989, 83 (01) :11-13

[6] Formation of de novo centromeres and construction of first-generation human artificial microchromosomes [J].

Harrington, JJ ;

VanBokkelen, G ;

Mays, RW ;

Gustashaw, K ;

Willard, HF .

NATURE GENETICS, 1997, 15 (04) :345-355

[7] Human artificial chromosomes generated by modification of a yeast artificial chromosome containing both human alpha satellite and single-copy DNA sequences [J].

Henning, KA ;

Novotny, EA ;

Compton, ST ;

Guan, XY ;

Liu, PP ;

Ashlock, MA .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (02) :592-597

[8] DISTRIBUTION OF CENP-B BOXES REFLECTED IN CREST CENTROMERE ANTIGENIC SITES ON LONG-RANGE ALPHA-SATELLITE DNA ARRAYS OF HUMAN-CHROMOSOME-21 [J].

IKENO, M ;

MASUMOTO, H ;

OKAZAKI, T .

HUMAN MOLECULAR GENETICS, 1994, 3 (08) :1245-1257

[9] Construction of YAC-based mammalian artificial chromosomes [J].

Ikeno, M ;

Grimes, B ;

Okazaki, T ;

Nakano, M ;

Saitoh, K ;

Hoshino, H ;

McGill, NI ;

Cooke, H ;

Masumoto, H .

NATURE BIOTECHNOLOGY, 1998, 16 (05) :431-439

[10] The case for epigenetic effects on centromere identity and function [J].

Karpen, GH ;

Allshire, RC .

TRENDS IN GENETICS, 1997, 13 (12) :489-496

← 1 2 →