Cargando…
Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization
Rats are effective model animals and have contributed to the development of human medicine and basic research. However, the application of reproductive engineering techniques to rats is not as advanced compared with mice, and genome editing in rats has not been achieved using embryos obtained by in...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6689013/ https://www.ncbi.nlm.nih.gov/pubmed/31399630 http://dx.doi.org/10.1038/s41598-019-47964-1 |
_version_ | 1783442968529600512 |
---|---|
author | Honda, Arata Tachibana, Ryoma Hamada, Kazuya Morita, Kohtaro Mizuno, Naoaki Morita, Kento Asano, Masahide |
author_facet | Honda, Arata Tachibana, Ryoma Hamada, Kazuya Morita, Kohtaro Mizuno, Naoaki Morita, Kento Asano, Masahide |
author_sort | Honda, Arata |
collection | PubMed |
description | Rats are effective model animals and have contributed to the development of human medicine and basic research. However, the application of reproductive engineering techniques to rats is not as advanced compared with mice, and genome editing in rats has not been achieved using embryos obtained by in vitro fertilization (IVF). In this study, we conducted superovulation, IVF, and knock out and knock in using IVF rat embryos. We found that superovulation effectively occurred in the synchronized oestrus cycle and with anti-inhibin antiserum treatment in immature rats, including the Brown Norway rat, which is a very difficult rat strain to superovulate. Next, we collected superovulated oocytes under anaesthesia, and offspring derived from IVF embryos were obtained from all of the rat strains that we examined. When the tyrosinase gene was targeted by electroporation in these embryos, both alleles were disrupted with 100% efficiency. Furthermore, we conducted long DNA fragment knock in using adeno-associated virus and found that the knock-in litter was obtained with high efficiency (33.3–47.4%). Thus, in this study, we developed methods to allow the simple and efficient production of model rats. |
format | Online Article Text |
id | pubmed-6689013 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-66890132019-08-13 Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization Honda, Arata Tachibana, Ryoma Hamada, Kazuya Morita, Kohtaro Mizuno, Naoaki Morita, Kento Asano, Masahide Sci Rep Article Rats are effective model animals and have contributed to the development of human medicine and basic research. However, the application of reproductive engineering techniques to rats is not as advanced compared with mice, and genome editing in rats has not been achieved using embryos obtained by in vitro fertilization (IVF). In this study, we conducted superovulation, IVF, and knock out and knock in using IVF rat embryos. We found that superovulation effectively occurred in the synchronized oestrus cycle and with anti-inhibin antiserum treatment in immature rats, including the Brown Norway rat, which is a very difficult rat strain to superovulate. Next, we collected superovulated oocytes under anaesthesia, and offspring derived from IVF embryos were obtained from all of the rat strains that we examined. When the tyrosinase gene was targeted by electroporation in these embryos, both alleles were disrupted with 100% efficiency. Furthermore, we conducted long DNA fragment knock in using adeno-associated virus and found that the knock-in litter was obtained with high efficiency (33.3–47.4%). Thus, in this study, we developed methods to allow the simple and efficient production of model rats. Nature Publishing Group UK 2019-08-09 /pmc/articles/PMC6689013/ /pubmed/31399630 http://dx.doi.org/10.1038/s41598-019-47964-1 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Honda, Arata Tachibana, Ryoma Hamada, Kazuya Morita, Kohtaro Mizuno, Naoaki Morita, Kento Asano, Masahide Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization |
title | Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization |
title_full | Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization |
title_fullStr | Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization |
title_full_unstemmed | Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization |
title_short | Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization |
title_sort | efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6689013/ https://www.ncbi.nlm.nih.gov/pubmed/31399630 http://dx.doi.org/10.1038/s41598-019-47964-1 |
work_keys_str_mv | AT hondaarata efficientderivationofknockoutandknockinratsusingembryosobtainedbyinvitrofertilization AT tachibanaryoma efficientderivationofknockoutandknockinratsusingembryosobtainedbyinvitrofertilization AT hamadakazuya efficientderivationofknockoutandknockinratsusingembryosobtainedbyinvitrofertilization AT moritakohtaro efficientderivationofknockoutandknockinratsusingembryosobtainedbyinvitrofertilization AT mizunonaoaki efficientderivationofknockoutandknockinratsusingembryosobtainedbyinvitrofertilization AT moritakento efficientderivationofknockoutandknockinratsusingembryosobtainedbyinvitrofertilization AT asanomasahide efficientderivationofknockoutandknockinratsusingembryosobtainedbyinvitrofertilization |