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Clark, Anthony; Burl, Sarah; Denning, Chris; Dickinson, Paul

doi:10.1023/a:1008974616402

Cited by 43 publications

(10 citation statements)

References 86 publications

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“…In conventional gene targeting, the targeting efficiency can be significantly improved by increasing the length of homologous arms. A longer arm of homology is more effective for searching the target locus and for spontaneous homologous recombination31. Consequently, we replaced the 3′ arm of pBLG-neo-M with a 5.3 kb homologous sequence to construct the targeting vector pBLG-neo (Fig.…”

Section: Resultsmentioning

confidence: 99%

Gene targeting by TALEN-induced homologous recombination in goats directs production of β-lactoglobulin-free, high-human lactoferrin milk

Cui

Song

et al. 2015

Sci Rep

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β-Lactoglobulin (BLG) is a major goat’s milk allergen that is absent in human milk. Engineered endonucleases, including transcription activator-like effector nucleases (TALENs) and zinc-finger nucleases, enable targeted genetic modification in livestock. In this study, TALEN-mediated gene knockout followed by gene knock-in were used to generate BLG knockout goats as mammary gland bioreactors for large-scale production of human lactoferrin (hLF). We introduced precise genetic modifications in the goat genome at frequencies of approximately 13.6% and 6.09% for the first and second sequential targeting, respectively, by using targeting vectors that underwent TALEN-induced homologous recombination (HR). Analysis of milk from the cloned goats revealed large-scale hLF expression or/and decreased BLG levels in milk from heterozygous goats as well as the absence of BLG in milk from homozygous goats. Furthermore, the TALEN-mediated targeting events in somatic cells can be transmitted through the germline after SCNT. Our result suggests that gene targeting via TALEN-induced HR may expedite the production of genetically engineered livestock for agriculture and biomedicine.

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Section: Resultsmentioning

confidence: 99%

Gene targeting by TALEN-induced homologous recombination in goats directs production of β-lactoglobulin-free, high-human lactoferrin milk

Cui

Song

et al. 2015

Sci Rep

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“…The genetic material from the donor cell is "reprogrammed" into totipotency by the recipient oocyte, such that the "reconstructed" egg is able to develop into a viable animal [7]. NT per se is effective in a very broad range of animal types including cattle, goats, mice and sheep [9]. Transgenes can be introduced to donor cells in vitro, permitting the production of genetically modified animals by NT [33].…”

Section: Cell Typesmentioning

confidence: 99%

Theoretical mechanisms in targeted and random integration of transgene DNA

Smith

2001

Reprod. Nutr. Dev.

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“…Unlike mice, in pigs genetic modifications need to be made in somatic cells that are then used for SCNT due to the lack of embryonic stem cells. However, the frequency of gene targeting in somatic cells is extremely low89. Recent developments in zinc finger nuclease (ZFN) technology allow us to modify the genome with precision and high efficiency.…”

mentioning

confidence: 99%

Production of biallelic CMP-Neu5Ac hydroxylase knock-out pigs

Kwon

Lee

Kang

et al. 2013

Sci Rep

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After the knock-out (KO) of α1,3 galactosyltransfease (Gal-T), the Hanganutziu-Deicher antigen became a major antigen of the "non-Gal antigen" that is implicated in subsequent xenograft rejection. For deletion of non-Gal antigen, we successfully produced zinc finger nuclease (ZFN)-mediated monoallelic/biallelic male and female CMP-N-acetylneuraminic acid hydroxylase (CMAH) KO miniature pigs: the efficiency of the gene targeting (41.7%) was higher when donor DNA was used with the ZFN than those of ZFN alone (9.1%). Monoallelic KO pigs had no integration of exogenous DNA into their genome, indicating that this technique would provide a new avenue to reduce the risk of antibiotics resistance when organs from genetically modified pigs are transplanted into patients. Until now, both monoallelic and biallelic CMAH KO pigs are healthy and show no sign of abnormality and off-target mutations. Therefore, these CMAH null pigs on the Gal-T KO background could serve as an important model for the xenotransplantation.

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Untitled

Cited by 43 publications

References 86 publications

Gene targeting by TALEN-induced homologous recombination in goats directs production of β-lactoglobulin-free, high-human lactoferrin milk

Gene targeting by TALEN-induced homologous recombination in goats directs production of β-lactoglobulin-free, high-human lactoferrin milk

Theoretical mechanisms in targeted and random integration of transgene DNA

Production of biallelic CMP-Neu5Ac hydroxylase knock-out pigs

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