Comparative Analysis of Genome Editors Efficiency on a Model of Mice Zygotes Microinjection

被引:3
作者
Averina, Olga A. [1 ,2 ]
Permyakov, Oleg A. [1 ]
Grigorieva, Olga O. [1 ]
Starshin, Alexey S. [3 ]
Mazur, Alexander M. [3 ]
Prokhortchouk, Egor B. [3 ]
Dontsova, Olga A. [4 ,5 ,6 ]
Sergiev, Petr, V [1 ,4 ,5 ]
机构
[1] Lomonosov Moscow State Univ, Inst Funct Genom, Moscow 119991, Russia
[2] Lomonosov Moscow State Univ, Belozersky Inst Physicochem Biol, Moscow 119899, Russia
[3] Russian Acad Sci, Inst Bioengn, Res Ctr Biotechnol, Moscow 119071, Russia
[4] Lomonosov Moscow State Univ, Dept Chem, Moscow 119991, Russia
[5] Skolkovo Inst Sci & Technol, Ctr Life Sci, Moscow 121205, Russia
[6] Shemyakin Ovchinnikov Inst Bioorgan Chem, Moscow 117997, Russia
基金
俄罗斯科学基金会;
关键词
genome editing; CRISPR Cas9; knockout mice; microinjection; homologous recombination; prime editor; MUTATIONS;
D O I
10.3390/ijms221910221
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Genome editing is an indispensable tool for functional genomics. The caveat of the genome-editing pipeline is a prevalence of error-prone non-homologous end joining over homologous recombination, while only the latter is suitable to introduce particularly desired genetic variants. To overcome this problem, a toolbox of genome engineering was appended by a variety of improved instruments. In this work, we compared the efficiency of a number of recently suggested improved systems for genome editing applied to the same genome regions on a murine zygote model via microinjection. As a result, we observed that homologous recombination utilizing an ssDNA template following sgRNA directed Cas9 cleavage is still the method of choice for the creation of animals with precise genome alterations.
引用
收藏
页数:10
相关论文
共 14 条
[11]   Production of a Cloned Offspring and CRISPR/Cas9 Genome Editing of Embryonic Fibroblasts in Cattle [J].
Singina, G. N. ;
Sergiev, P., V ;
Lopukhov, A., V ;
Rubtsova, M. P. ;
Taradajnic, N. P. ;
Ravin, N., V ;
Shedova, E. N. ;
Taradajnic, T. E. ;
Polejaeva, I. A. ;
Dozev, A., V ;
Brem, G. ;
Dontsova, O. A. ;
Zinovieva, N. A. .
DOKLADY BIOCHEMISTRY AND BIOPHYSICS, 2021, 496 (01) :48-51
[12]   Stem Cells, Genome Editing, and the Path to Translational Medicine [J].
Soldner, Frank ;
Jaenisch, Rudolf .
CELL, 2018, 175 (03) :615-632
[13]   Mutation of C20orf7 Disrupts Complex I Assembly and Causes Lethal Neonatal Mitochondrial Disease [J].
Sugiana, Canny ;
Pagliarini, David J. ;
McKenzie, Matthew ;
Kirby, Denise M. ;
Salemi, Renato ;
Abu-Amero, Khaled K. ;
Dahl, Hans-Henrik M. ;
Hutchison, Wendy M. ;
Vascotto, Katherine A. ;
Smith, Stacey M. ;
Newbold, Robert F. ;
Christodoulou, John ;
Calvo, Sarah ;
Mootha, Vamsi K. ;
Ryan, Michael T. ;
Thorburn, David R. .
AMERICAN JOURNAL OF HUMAN GENETICS, 2008, 83 (04) :468-478
[14]   One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering [J].
Wang, Haoyi ;
Yang, Hui ;
Shivalila, Chikdu S. ;
Dawlaty, Meelad M. ;
Cheng, Albert W. ;
Zhang, Feng ;
Jaenisch, Rudolf .
CELL, 2013, 153 (04) :910-918