Lanh Thi Kim Do
FVM- Vietnam National University of Agriculture
Keywords: clustered regularly interspaced short palindromic repeats/Cas9, electroporation, gene editing, microinjection, porcine zygotes. LinkPublications, Science and Technology
Triple gene editing in porcine embryos using electroporation alone or in combination with microinjection
Abstract
Background and Aim: We previously developed the gene-editing by electroporation (EP) of Cas9 protein method, in which the CRISPR/Cas9 system was introduced into porcine in vitro fertilized (IVF) zygotes through EP to disrupt a target gene. This method should be further developed, and a combination of EP and MI methods should be evaluated in pigs. This study aimed to determine that a combination of microinjection (MI) and EP of CRISPR/Cas9 system could increase the rates of biallelic mutation for triple-gene knockout in porcine blastocysts. We targeted the pancreatic and duodenal homeobox1 (PDX1) gene using cytoplasmic MI 1 h before or after EP, which was used to edit alpha-1,3-galactosyltransferase (GGTA1) and cytidine 32 monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) genes in porcine zygotes.
Materials and Methods: We introduced guide RNAs targeting PDX1, GGTA1, and CMAH with the Cas9 protein into IVF zygotes (one-cell stage) through EP 10 h after the start of IVF (IVF; EP group) or in combination with MI (1 h before, MI-EP group, or after EP treatment EP-MI group) and evaluated the blastocyst formation rate and efficiency of target mutations in the resulting blastocysts.
Results: Our results revealed a significant reduction in the rate of blastocyst formation in the two groups that underwent MI before and after EP (MI-EP and EP-MI group), compared with that in the groups treated with EP alone (EP group) (p=0.0224 and p<0.0001, respectively) and control (p=0.0029 and p<0.0001, respectively). There was no significant difference in the total mutation rates among the treatment groups in the resulting blastocysts. As an only positive effect of additional MI treatment, the rate of blastocysts carrying biallelic mutations in at least one target gene was higher in the MI-EP group than in the EP group. However, there was no difference in the rates of embryos carrying biallelic mutations in more than 2 target genes.
Conclusion: These results indicate that although a combination of MI and EP does not improve the mutation efficiency or biallelic mutation for triple-gene knockout, MI treatment before EP is better to reduce mortality in porcine zygotic gene editing through a combination of MI and EP.
Tiếng Việt
I like it when people get together and share thoughts. Great blog, keep it up!
Kender du nogen metoder, der kan hjælpe med at forhindre, at indholdet bliver stjålet? Det ville jeg sætte stor pris på.
buď vytvořil sám, nebo zadal externí firmě, ale vypadá to.
Hello! This is my 1st comment here so I just wanted to give a quick shout out and say I truly enjoy reading through your blog posts. Can you recommend any other blogs/websites/forums that deal with the same topics? Thanks a ton!
) Znovu ho navštívím, protože jsem si ho poznamenal. Peníze a svoboda je nejlepší způsob, jak se změnit, ať jste bohatí a
reading this weblog’s post to be updated daily.
for the reason that here every material is quality based
O conteúdo existente nesta página é realmente notável para a experiência das pessoas,
Conhecem algum método para ajudar a evitar que o conteúdo seja roubado? Agradecia imenso.
Fiquei muito feliz em descobrir este site. Preciso de agradecer pelo vosso tempo
Também tenho o seu livro marcado para ver coisas novas no seu blog.
e dizer que gosto muito de ler os vossos blogues.
Obrigado|Olá a todos, os conteúdos existentes nesta
مرحبًا، أعتقد أن هذه مدونة ممتازة. لقد عثرت عليها بالصدفة ;
Kan du anbefale andre blogs / websteder / fora, der beskæftiger sig med de samme emner?
pokračujte v pěkné práci, kolegové.|Když máte tolik obsahu a článků, děláte to?
Tak Hej der til alle, det indhold, der findes på denne
webside er virkelig bemærkelsesværdig for folks oplevelse, godt,
že spousta z něj se objevuje na internetu bez mého souhlasu.
الاستمرار في توجيه الآخرين.|Ahoj, věřím, že je to vynikající blog. Narazil jsem na něj;
Esta página tem definitivamente toda a informação que eu queria sobre este assunto e não sabia a quem perguntar. Este é o meu primeiro comentário aqui, então eu só queria dar um rápido
) Znovu ho navštívím, protože jsem si ho poznamenal. Peníze a svoboda je nejlepší způsob, jak se změnit, ať jste bohatí a
nogensinde løbe ind i problemer med plagorisme eller krænkelse af ophavsretten? Mit websted har en masse unikt indhold, jeg har
Díky moc!|Hej, jeg synes, dette er en fremragende blog. Jeg snublede over det;
det. Denne side har bestemt alle de oplysninger, jeg ønskede om dette emne, og vidste ikke, hvem jeg skulle spørge. Dette er min 1. kommentar her, så jeg ville bare give en hurtig
že spousta z něj se objevuje na internetu bez mého souhlasu.
nenarazili jste někdy na problémy s plagorismem nebo porušováním autorských práv? Moje webové stránky mají spoustu unikátního obsahu, který jsem vytvořil.
muito dele está a aparecer em toda a Internet sem o meu acordo.
Obrigado|Olá a todos, os conteúdos existentes nesta
také jsem si vás poznamenal, abych se podíval na nové věci na vašem blogu.|Hej! Vadilo by vám, kdybych sdílel váš blog s mým facebookem.
It contains fastidious material.|I think the admin of this website is actually working hard in favor of his site,
webside er virkelig bemærkelsesværdig for folks oplevelse, godt,
har også bogmærket dig for at se på nye ting på din blog Hej! Har du noget imod, hvis jeg deler din blog med min facebook
Esta página tem definitivamente toda a informação que eu queria sobre este assunto e não sabia a quem perguntar. Este é o meu primeiro comentário aqui, então eu só queria dar um rápido
Com tanto conteúdo e artigos, vocês já se depararam com algum problema de plágio?
skupině? Je tu spousta lidí, o kterých si myslím, že by se opravdu
Does your blog have a contact page? I’m having problems locating it but, I’d like to send you an email. I’ve got some ideas for your blog you might be interested in hearing. Either way, great website and I look forward to seeing it improve over time.
for the reason that here every material is quality based
ocenili váš obsah. Dejte mi prosím vědět.
devido a esta maravilhosa leitura!!! O que é que eu acho?
After going over a few of the blog posts on your web page, I honestly appreciate your way of blogging. I bookmarked it to my bookmark website list and will be checking back soon. Please check out my web site too and let me know what you think. https://www.sheffieldstateuniversity.com/
ocenili váš obsah. Dejte mi prosím vědět.
Tak Hej der til alle, det indhold, der findes på denne
This is really interesting, You are a very skilled blogger. I’ve joined your rss feed and look forward to seeking more of your excellent post. Also, I’ve shared your site in my social networks!
|Tato stránka má rozhodně všechny informace, které jsem o tomto tématu chtěl a nevěděl jsem, koho se zeptat.|Dobrý den! Tohle je můj 1. komentář tady, takže jsem chtěl jen dát rychlý
Hmm it seems like your website ate my first comment (it was extremely long) so I guess I’ll just sum it up what I submitted and say, I’m thoroughly enjoying your blog. I as well am an aspiring blog blogger but I’m still new to everything. Do you have any tips for rookie blog writers? I’d really appreciate it.
Muito obrigado!}
pokračujte v pěkné práci, kolegové.|Když máte tolik obsahu a článků, děláte to?
information.|My family members every time say that I am killing my time here
It contains fastidious material.|I think the admin of this website is actually working hard in favor of his site,
It contains fastidious material.|I think the admin of this website is actually working hard in favor of his site,
har også bogmærket dig for at se på nye ting på din blog Hej! Har du noget imod, hvis jeg deler din blog med min facebook
fortsæt det gode arbejde stipendiater. Med at have så meget indhold og artikler gør du det
Com tanto conteúdo e artigos, vocês já se depararam com algum problema de plágio?
at web, except I know I am getting familiarity all the time by reading thes pleasant posts.|Fantastic post. I will also be handling some of these problems.|Hello, I think this is a great blog. I happened onto it;) I have bookmarked it and will check it out again. The best way to change is via wealth and independence. May you prosper and never stop mentoring others.|I was overjoyed to find this website. I must express my gratitude for your time because this was an amazing read! I thoroughly enjoyed reading it, and I’ve bookmarked your blog so I can check out fresh content in the future.|Hi there! If I shared your blog with my Facebook group, would that be okay? I believe there are a lot of people who would truly value your article.|منشور رائع. سأتعامل مع بعض هذه|
Podem recomendar outros blogues/sites/fóruns que tratem dos mesmos temas?
Hello! Someone in my Facebook group shared this site with us so I came to give it a look. I’m definitely enjoying the information. I’m book-marking and will be tweeting this to my followers! Fantastic blog and excellent style and design.
enten oprettet mig selv eller outsourcet, men det ser ud til
Wonderful blog! I found it while searching on Yahoo News. Do you have any suggestions on how to get listed in Yahoo News? I’ve been trying for a while but I never seem to get there! Thank you
på grund af denne vidunderlige læsning !!! Jeg kunne bestemt virkelig godt lide hver eneste lille smule af det, og jeg
Muito obrigado!}
fortsæt med at guide andre. Jeg var meget glad for at afdække dette websted. Jeg er nødt til at takke dig for din tid
Díky moc!|Hej, jeg synes, dette er en fremragende blog. Jeg snublede over det;
muito dele está a aparecer em toda a Internet sem o meu acordo.
grupo do facebook? Há muitas pessoas que eu acho que iriam realmente
fortsæt det gode arbejde stipendiater. Med at have så meget indhold og artikler gør du det
Everything is very open with a really clear clarification of the challenges. It was really informative. Your site is extremely helpful. Many thanks for sharing.
Thanks for the post, can I set it up so I get an update sent in an email every time there is a new article?
Com tanto conteúdo e artigos, vocês já se depararam com algum problema de plágio?
The Natural Mounjaro Recipe is more than just a diet—it’s a sustainable and natural approach to weight management and overall health.
There is certainly a lot to learn about this issue. I really like all of the points you have made.