Site-specific chromosomal integration in mammalian cells: highly efficient CRE recombinase-mediated cassette exchange

Feng, Yong; Seibler, Jost; Alami, Ramzi S.; Eisen, Andrew; Westerman, Karen A.; Leboulch, Philippe; Fiering, Steven; Bouhassira, Eric E.

doi:10.1006/jmbi.1999.3113

Cited by 191 publications

(219 citation statements)

References 16 publications

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“…Consistent with this hypothesis is the finding that even the prototype for locus control regions, the ␤-like globin locus control region, is affected by the site of integration and does not ensure expression levels equal to those of the endogenous locus (47)(48)(49). This has been made even more apparent by studies in which the same gene is repeatedly integrated into several, distinct chromosomal sites by CRE recombinase-mediated cassette exchange (50,51).…”

Section: Discussionmentioning

confidence: 91%

Transcription of a Productively Rearranged Ig VDJCα Does Not Require the Presence of HS4 in the Igh 3′ Regulatory Region

Zhang

Alaie-Petrillo

Kon

et al. 2007

The Journal of Immunology

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V gene assembly, class switch recombination, and somatic hypermutation are gene-modifying processes essential to the development of an effective Ab response. If inappropriately applied, however, these processes can mediate genetic changes that lead to disease (e.g., lymphoma). A series of control elements within the Ig H chain (Igh) locus has been implicated in regulating these processes as well as in regulating IgH gene transcription. These include the intronic enhancer (Eμ) and several elements at the 3′ end of the locus (hs1,2, hs3a, hs3b, and hs4) known collectively as the 3′ regulatory region. Although it is clear that the Eμ plays a unique role in V gene assembly, it has not been established whether there are unique functions for each element within the 3′ regulatory region. In earlier studies in mice and in mouse cell lines, pairwise deletion of hs3b and hs4 had a dramatic effect on both class switch recombination and IgH gene transcription; deletion of an element almost identical with hs3b (hs3a), however, yielded no discernible phenotype. To test the resulting hypothesis that hs4 is uniquely required for these processes, we induced the deletion of hs4 within a bacterial artificial chromosome transgene designed to closely approximate the 3′ end of the natural Igh locus. When introduced into an Ig-secreting cell line, an Igα transcription unit within the bacterial artificial chromosome was expressed efficiently and the subsequent deletion of hs4 only moderately affected Igα expression. Thus, hs4 does not play a uniquely essential role in the transcription of a productively rearranged Ig VDJCα transcription unit.

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Section: Discussionmentioning

confidence: 91%

Transcription of a Productively Rearranged Ig VDJCα Does Not Require the Presence of HS4 in the Igh 3′ Regulatory Region

Zhang

Alaie-Petrillo

Kon

et al. 2007

The Journal of Immunology

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“…While mutations might accumulate with time, AID will eventually get inactivated for mutating itself so the likelihood of a disabling mutation in the V region of switch variants is low. Nevertheless, this low potential for non desired mutations can be prevented by removing the AID gene from switched cells for example by using a Cre-lox strategy (Feng et al, 1999).…”

Section: Discussionmentioning

confidence: 99%

Forced expression of AID facilitates the isolation of class switch variants from hybridoma cells

Iglesias-Ussel

Fan

et al. 2006

Journal of Immunological Methods

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Monoclonal antibodies are used in the treatment and diagnosis of diseases and to study the protective and adverse functions of antibodies in vitro and in vivo. Since the isotype determines the effector function, half-life in the serum and distribution throughout the body, it would be useful to have a battery of antibodies with the same binding site associated with different isotypes. However, since hybridomas switch isotypes at very low frequencies in tissue culture, it has been difficult and very labor intensive to isolate panels of class switch variants. We show here that stable transfection of activation-induced cytidine deaminase (AID) in hybridomas increased their frequency of switching to a level that greatly facilitated the isolation of subclones expressing monoclonal antibodies of different isotypes. Although forced expression of AID also increased the frequency of somatic hypermutation in the immunoglobulin variable regions that encode the antigen-binding site, antigen recognition was retained in the isotype switched antibodies.

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“…The use of two independent and noncompatible LoxP sites instead of one in the vectors and one in the site of integration appears presently to be the most attractive way of using the LoxP-Cre system to integrate a foreign gene in a targeted region of the genome [3,10].…”

Section: Gene Targetingmentioning

confidence: 99%

Transgenesis for the study and the control of lactation

Houdebiné

Rival

Pantano³

et al. 2002

Reprod. Nutr. Dev.

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-The study and the control of milk synthesis are required to decipher the mechanisms of gene expression, to improve milk production, to modify milk composition, to induce a resistance to diseases in the mammary gland and to produce recombinant proteins of pharmaceutical interest. Transgenesis has become a mandatory tool to reach these goals. The use of transgenesis is still limited by the difficulty of adding foreign genes in farm animals and mainly by replacing genes by homologous recombination. Transgene expression is also often ill-controlled. The present paper summarizes the current progress in this field with a particular emphasis on expression vectors for transgenes. lactation / transgenesis / expression vectorsRésumé -La transgénèse pour l'étude et le contrôle de la lactation. L'étude et le contrôle de la synthèse du lait sont nécessaires pour décrypter les mécanismes de l'expression de gènes, pour amé-liorer la production de lait, pour modifier la composition du lait, pour induire une résistance de la glande mammaire contre des maladies infectieuses et pour produire des protéines recombinantes d'intérêt pharmaceutique. La transgénèse est devenue un outil indispensable pour mener à bien ces projets. La mise en oeuvre de la transgénèse est encore limitée par la difficulté d'ajouter des gènes étrangers chez les animaux d'élevage et surtout de remplacer des gènes par recombinaison homologue. L'expression des transgènes est par ailleurs souvent mal contrôlée. Cet article se propose de faire le point dans ce domaine en développant plus particulièrement la mise au point de vecteurs d'expression pour les transgènes. lactation / transgénèse / vecteurs d'expression

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Site-specific chromosomal integration in mammalian cells: highly efficient CRE recombinase-mediated cassette exchange

Cited by 191 publications

References 16 publications

Transcription of a Productively Rearranged Ig VDJCα Does Not Require the Presence of HS4 in the Igh 3′ Regulatory Region

Transcription of a Productively Rearranged Ig VDJCα Does Not Require the Presence of HS4 in the Igh 3′ Regulatory Region

Forced expression of AID facilitates the isolation of class switch variants from hybridoma cells

Transgenesis for the study and the control of lactation

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