Cattle with a precise, zygote-mediated deletion safely eliminate the major milk allergen beta-lactoglobulin

Scientific Reports
Jingwei WeiGötz Laible

Abstract

We applied precise  zygote-mediated genome editing to eliminate beta-lactoglobulin (BLG), a major allergen in cows' milk. To efficiently generate LGB knockout cows, biopsied embryos were screened to transfer only appropriately modified embryos. Transfer of 13 pre-selected embryos into surrogate cows resulted in the birth of three calves, one dying shortly after birth. Deep sequencing results confirmed conversion of the genotype from wild type to the edited nine bp deletion by more than 97% in the two male calves. The third calf, a healthy female, had in addition to the expected nine bp deletion (81%), alleles with an in frame 21 bp deletion (<17%) at the target site. While her milk was free of any mature BLG, we detected low levels of a BLG variant derived from the minor deletion allele. This confirmed that the nine bp deletion genotype completely knocks out production of BLG. In addition, we showed that the LGB knockout animals are free of any TALEN-mediated off-target mutations or vector integration events using an unbiased whole genome analysis. Our study demonstrates the feasibility of generating precisely biallelically edited cattle by zygote-mediated editing for the safe production of hypoallergenic milk.

References

Dec 19, 2002·Annals of Allergy, Asthma & Immunology : Official Publication of the American College of Allergy, Asthma, & Immunology·Arne Høst
Jan 28, 2003·Nature Biotechnology·Brigid BrophyGötz Laible
May 27, 2008·Nature Biotechnology·Yannick DoyonSharon L Amacher
May 27, 2008·Nature Biotechnology·Xiangdong MengScot A Wolfe
Jun 10, 2009·Bioinformatics·Heng LiUNKNOWN 1000 Genome Project Data Processing Subgroup
Jul 25, 2009·Science·Aron M GeurtsRoland Buelow
Oct 3, 2012·Proceedings of the National Academy of Sciences of the United States of America·Anower JabedGoetz Laible
Jan 1, 2013·The Journal of Allergy and Clinical Immunology·Robert A WoodHugh A Sampson
Apr 30, 2013·Dairy Science & Technology·Guanhao BuTingwei Zhu
Jun 25, 2013·Nature Biotechnology·Yanfang FuJeffry D Sander
Aug 14, 2013·Nucleic Acids Research·Thomas J CradickGang Bao
Sep 5, 2014·PloS One·Wei NiChuangfu Chen
Oct 11, 2014·Nucleic Acids Research·Eva K BrinkmanBas van Steensel
Mar 25, 2015·Allergologia et immunopathologia·A Martorell-AragonésUNKNOWN Food allergy committee of SEICAP (Spanish Society of Pediatric Allergy, Asthma and Clinical Immunology)
May 29, 2015·Nature Methods·Vivek IyerWilliam C Skarnes
Sep 27, 2015·Veterinary Research·Grant A SmolenskiThomas T Wheeler
Feb 10, 2016·Scientific Reports·Xianlong WangJianguo Zhao
Feb 24, 2016·Scientific Reports·Simon G LillicoC Bruce A Whitelaw
May 8, 2016·Nature Biotechnology·Daniel F CarlsonScott C Fahrenkrug
Apr 30, 2017·Methods in Molecular Biology·Henrik Nielsen
Jun 21, 2017·The FEBS Journal·Wen-Rong LiMing-Jun Liu
Oct 11, 2017·PloS One·Wenjun ZhouFeng Wang

❮ Previous
Next ❯

Citations

Sep 28, 2018·Frontiers in Plant Science·Steffi FritscheGlenn Thorlby
Oct 9, 2019·Nature Biotechnology·Amy E YoungAlison L Van Eenennaam
Oct 1, 2020·Scientific Reports·Joseph R OwenAlison L Van Eenennaam
Feb 9, 2020·The Journal of Experimental Biology·Thomas Frederick Bishop, Alison Louise Van Eenennaam
Dec 1, 2020·Molecular Reproduction and Development·Zachariah L McLeanBjörn Oback
Jan 15, 2019·Molecular Therapy. Nucleic Acids·Subbroto Kumar SahaKi-Hyun Kim
Mar 23, 2021·Frontiers in Genetics·Laura Daniela RatnerRafael Fernandez-Martin
Jun 8, 2021·Frontiers in Physiology·Lindsay Sawyer

❮ Previous
Next ❯

Methods Mentioned

BETA
genetic modifications
gene exchange
biopsy
PCR
biopsies
whole genome shotgun sequencing
transgenic
electrophoresis
genotyping
FCS

Software Mentioned

Perl script
SignalP
Quantity One
GATK IndelRealigner
TIDE calculator
BWA
PROGNOS
PEAKS Studio
SPIDER
[UNK] Analysis Pro

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.