Site-specific integrase-mediated transgenesis in mice via pronuclear injection

Tasic, Bosiljka; Hippenmeyer, Simon; Wang, Charlene; Gamboa, Matthew; Zong, Hui; Chen-Tsai, Yanru; Luo, Liqun

doi:10.1073/pnas.1019507108

Cited by 215 publications

(269 citation statements)

References 48 publications

(55 reference statements)

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“…These reporter constructs are either injected into embryos to generate stable transformants (Venken and Bellen, 2007;Tasic et al, 2011;Frokjaer-Jensen et al, 2012), or electroporated (Funahashi and Nakamura, 2008;Vierra and Irvine, 2012), to observe the transient effect of the construct. In most cases, these small DNA fragments are sufficient to provide coherent functions in vivo.…”

Section: Interactions Between Regulatory Elementsmentioning

confidence: 99%

Multiple layers of complexity in cis‐regulatory regions of developmental genes

Frankel

2012

Developmental Dynamics

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Genomes contain the necessary information to ensure that genes are expressed in the right place, at the right time, and with the proper rate. Metazoan developmental genes often possess long stretches of DNA flanking their coding sequences and/or large introns which contain elements that influence gene expression. Most of these regulatory elements are relatively small and can be studied in isolation. For example, transcriptional enhancers, the elements that generate the expression pattern of a gene, have been traditionally studied with reporter constructs in transgenic animals. These studies have provided and will provide invaluable insights into enhancer evolution and function. However, this experimental approach has its limits; often, enhancer elements do not faithfully recapitulate native expression patterns. This fact suggests that additional information in cis-regulatory regions modulates the activity of enhancers and other regulatory elements. Indeed, recent studies have revealed novel functional aspects at the level of whole cis-regulatory regions. First, the discovery of ''shadow enhancers.'' Second, the ubiquitous interactions between cis-regulatory elements. Third, the notion that some cis-regulatory regions may not function in a modular manner. Last, the effect of chromatin conformation on cis-regulatory activity. In this article, I describe these recent findings and discuss open questions in the field.

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Section: Interactions Between Regulatory Elementsmentioning

confidence: 99%

Multiple layers of complexity in cis‐regulatory regions of developmental genes

Frankel

2012

Developmental Dynamics

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“…2B). As a result, they obtained correctly integrated embryos and pups with high efficiencies (3.0-40.0%) ( Table 1) [67]. the random integration rate, however, was also high (0-20.0%) compared with the Cre-PITT that we reported.…”

Section: Phic31 Integrase-based Pitt (Phic31-pitt)mentioning

confidence: 67%

“…The PhiC31 integrase system for targeted integration through pronuclear injection in mice was recently reported by tasic et al [67]. They used a single full-length attP or three shortened tandem attP sites [27] and placed them into the Rosa26 and Hipp11 (H11) loci.…”

Section: Phic31 Integrase-based Pitt (Phic31-pitt)mentioning

confidence: 99%

“…tasic et al [67] utilized the H11 locus in the PhiC31-based PITT system because they expected that H11 might allow better access to PhiC31 integrase than Rosa26 based on their previous study using MaDM cassettes (containing CAG promoter-driven transgenes, loxP, and FRT sites) integrated into the H11 locus, which showed a high level of global transgene expression and a higher rate of interchromosomal recombination compared with those integrated into the Rosa26 locus [30,67]. In fact tasic et al successfully integrated a single-copy transgene into the H11 locus using a PhiC31-PITT system with high efficiency (up to 40%) [67].…”

Section: Hipp11 (H11)mentioning

confidence: 99%

“…In all of the PITT-related technologies, the Rosa26 locus has been the first choice to test the feasibility of the method [44,46,47,67], and in fact, targeted Tg mice at this locus have been readily obtained with these methods. thus, the Rosa26 locus can confer efficient targeted integration of DOI in ES cells as well as zygotes.…”

Section: Rosa26mentioning

confidence: 99%

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PITT: Pronuclear Injection-Based Targeted Transgenesis, a Reliable Transgene Expression Method in Mice

et al. 2012

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Transgenic (Tg) mice have been extensively used as valuable tools for analyses of gene function and have also served as models for many human diseases. Typically, a transgenic mouse is created by microinjection of DNA into pronuclei in which the DNA gets integrated at random locations in the genome. Frequently however, the random integration of multiple copies of a transgene results in transgene silencing, probably because of a positional effect and/or repeat-induced gene silencing. The transgene silencing issue has been overcome by single-copy transgene integration into a predetermined locus through ES cell-mediated transgenesis, despite it being expensive and more time-consuming compared with pronuclear injection (PI)-mediated transgenesis. Recently, several groups have reported novel approaches that employ PI for targeted transgenesis. They are based on site-specific recombination catalyzed by a recombinase or an integrase or homologous recombination enhanced by a zinc-finger nuclease via PI. These next-generation transgenesis methods, which we termed as PI-based Targeted Transgenesis (PITT), are more convenient and faster than ES cell-based transgenesis. Furthermore, the Tg mice generated by these newer methods contain a single-copy transgene and exhibit reliable expression of the transgene. The objective of this review is to present the recent progress in mouse targeted transgenesis.

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A Review of Single‐Cell Pose Adjustment and Puncture

Guo

Zhang

Jin

et al. 2022

Advanced Intelligent Systems

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Single‐cell manipulation technology is an essential method of cell research, with wide applications in biological engineering, medical engineering, agricultural engineering, and other precise manipulation fields. Cell pose adjustment and cell puncture are considered as two basic operations of single‐cell manipulation. Cell pose adjustment can be used for cell sorting, cell trapping, cell orientation, cell transportation, etc. It consists of direct contact manipulation methods and noncontact manipulation methods (e.g., mechanical contact method, electric field, optical field, magnetic field, acoustic field, and hydrodynamic methods). Cell puncture consists of ordinary glass needle puncture methods, piezoelectric‐assisted puncture methods, and rotational oscillatory drill puncture methods. It aims to achieve directional and quantified injection or sampling with minimal damage. Herein, the recent research progress on single‐cell pose adjustment and puncture methods is systematically summarized and discussed, which involves the basic mechanism, characteristics, limitations, and applications. Some potential directions for future research are proposed in accordance with the above analysis. It is hoped that this review can provide a reference for academic research and industrial applications of single‐cell manipulation technology.

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Site-specific integrase-mediated transgenesis in mice via pronuclear injection

Cited by 215 publications

References 48 publications

Multiple layers of complexity in cis‐regulatory regions of developmental genes

Multiple layers of complexity in cis‐regulatory regions of developmental genes

PITT: Pronuclear Injection-Based Targeted Transgenesis, a Reliable Transgene Expression Method in Mice

A Review of Single‐Cell Pose Adjustment and Puncture

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