scholarly journals Pronuclear Injection‐Based Targeted Transgenesis

2016 ◽  
Vol 91 (1) ◽  
Author(s):  
Samantha L.P. Schilit ◽  
Masato Ohtsuka ◽  
Rolen M. Quadros ◽  
Channabasavaiah B. Gurumurthy
Keyword(s):  
Pronuclear Injection

Production of YAC Transgenic Mice by Pronuclear Injection

2002 ◽  
pp. 55-65
Author(s):  
Justin F.-X. Ainscough ◽  
Rosalind M. John ◽  
Sheila C. Barton
Keyword(s):  
Transgenic Mice ◽  
Pronuclear Injection

Generation ofThy1Constructs for Pronuclear Injection

2015 ◽  
Vol 2015 (10) ◽  
pp. pdb.prot087676
Author(s):  
Petar Marinković ◽  
Leanne Godinho ◽  
Thomas Misgeld
Keyword(s):  
Pronuclear Injection

Genetransfer by pronuclear injection in the bovine

1988 ◽  
Vol 29 (1) ◽  
pp. 224 ◽  
Author(s):  
K.A. Biery ◽  
K.R. Bondioli ◽  
F.J. De Mayo
Keyword(s):  
Pronuclear Injection

scholarly journals Highly-efficient Cpf1-mediated gene targeting in mice following high concentration pronuclear injection

2016 ◽  
Author(s):  
Dawn E. Watkins-Chow ◽  
Gaurav K. Varshney ◽  
Lisa J. Garrett ◽  
Zelin Chen ◽  
Erin A. Jimenez ◽  
...  
Keyword(s):  
Gene Targeting ◽  
High Concentration ◽  
Highly Efficient ◽  
Pronuclear Injection

ABSTRACTCpf1 has emerged as an alternative to the Cas9 RNA-guided nuclease. Here we show that gene targeting rates in mice using Cpf1 can meet or even surpass Cas9 targeting rates (approaching 100% targeting) but require higher concentrations of mRNA and guide. We also demonstrate that co-injecting two guides with close targeting sites can result in synergistic genomic cutting, even if one of the guides has minimal cutting activity.

scholarly journals An optimized electroporation approach for efficient CRISPR/Cas9 genome editing in murine zygotes

2018 ◽  
Author(s):  
Simon E. Tröder ◽  
Lena K. Ebert ◽  
Linus Butt ◽  
Sonja Assenmacher ◽  
Bernhard Schermer ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Genome Editing ◽  
Embryo Development ◽  
Technical Requirement ◽  
Embryo Viability ◽  
Physical Damage ◽  
Novel Approach ◽  
Pronuclear Injection ◽  
Pre Treatment ◽  
Injection Procedure

AbstractElectroporation of zygotes represents a rapid alternative to the elaborate pronuclear injection procedure for CRISPR/Cas9-mediated genome editing in mice. However, current protocols for electroporation either require the investment in specialized electroporators or corrosive pre-treatment of zygotes which compromises embryo viability. Here, we describe an easily adaptable approach for the introduction of specific mutations in C57BL/6N mice by electroporation of intact zygotes using a common electroporator with synthetic CRISPR/Cas9 components and minimal technical requirement. Direct comparison to conventional pronuclear injection demonstrates significantly reduced physical damage and thus improved embryo development with successful genome editing in up to 100% of living offspring. Hence, our novel approach for Easy Electroporation of Zygotes (EEZy) allows highly efficient generation of CRISPR/Cas9 transgenic mice while reducing the numbers of animals required.

Gene transfer in animal production

1988 ◽  
Vol 12 ◽  
pp. 1-14
Author(s):  
A.J. Clark
Keyword(s):  
Gene Expression ◽  
Gene Transfer ◽  
Molecular Level ◽  
Milk Composition ◽  
Farm Animals ◽  
Research Groups ◽  
Genetic Changes ◽  
Pharmaceutical Proteins ◽  
Pronuclear Injection ◽  
Recent Developments

ABSTRACTGene transfer by pronuclear injection has been accomplished in farm animals by a number of research groups. Applications of this technology for improving milk composition, producing pharmaceutical proteins and manipulating physiology are described. Recent developments in our understanding of gene expression at the molecular level will increase the precision with which genetic changes can be made by gene transfer.

scholarly journals A quantitative test for developmental neutrality of a transgenic lineage marker in mouse chimaeras

1996 ◽  
Vol 67 (2) ◽  
pp. 135-146 ◽  
Author(s):  
John D. West ◽  
Margaret A. Keighren ◽  
Jean H. Flockhart
Keyword(s):  
Globin Gene ◽  
Genetic Effect ◽  
Cell Marker ◽  
Developmental Processes ◽  
Glucose Phosphate ◽  
Pronuclear Injection ◽  
Transgenic Cells ◽  
Eye Pigmentation ◽  
Extraembryonic Membranes

SummaryThe mouse transgene, provisionally designated TgN(Hbb-b1)83Clo, was produced by Dr C. Lo by pronuclear injection of the cloned ß-major globin gene and comprises a highly reiterated sequence that is readily detected by DNA in situ hybridization on histological sections. This fulfils many of the requirements of an ideal genetic cell marker and has been widely used for lineage studies with mouse chimaeras. However, it is not known whether it causes cell selection or influences developmental processes, such as cell mixing, in chimaeric tissues. In the present study, nontransgenic genetic markers (electrophoretic polymorphisms of glucose phosphate isomerase and differences in eye pigmentation) revealed no significant effect of the presence of hemizygous transgenic cells on the overall composition, size or gross morphology of 12½ d chimaeric foetuses, placentas or extraembryonic membranes. Also, a previously described maternal genetic effect on the composition of chimaeric tissues occurred in the presence or absence of the transgene. These tests have demonstrated that hemizygous cells are not at a significant selective disadvantage, when incorporated into mouse aggregationchimaeras with non-transgenic cells. Further studies are needed to test whether homozygous transgenic cells are also selectively neutral and to test whether hemizygous or homozygous transgenic cells influence developmental processes, such as cell mixing, that were not tested.

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