|
- 积分
- 23
- 威望
- 23
- 包包
- 86
|
本帖最后由 zwb96 于 2013-5-9 23:41 编辑
2 x/ m6 Z# n$ D# N# J- \- N% r3 Z+ O. e1 K" f9 ]9 @, L2 b' k
刚刚在Cell发表的文章（2013.5.9），转基因小鼠技术的鼻祖Rudolf Jaenisch领导的小组采用继ZFN,TALEN技术后最新的可修饰哺乳动物基因组的技术CRISPR以不可思议的高效率在小鼠胚胎干细胞一步敲除多个基因，更在短时间内通过合子内注射CRISPR系统（Cas9蛋白和sg-RNAs)建立了双基因敲除的小鼠模型，理论上只需4周时间就可完成，对基因敲除的技术来说是一次很大的飞跃，因为采用传统的同源重组的方法一般要9-15个月才能得到单个基因敲除的小鼠，敲除两个基因需要两年甚至更长的时间。以下是文章摘要。
* @8 D; Q! K& L) d6 ~Mice carrying mutations in multiple genes are traditionally" r: S5 i! z% d7 g0 \3 A, ~
generated by sequential recombination in
4 ~! H, U+ h8 m8 Z3 kembryonic stem cells and/or time-consuming intercrossing
& _" R! ~# n$ M) Mof mice with a single mutation. The
# a2 H* L. {- T' dCRISPR/Cas system has been adapted as an efficient8 U/ ?: m" x' t$ b* {
gene-targeting technology with the potential
5 e' q7 K7 B- H4 a3 rfor multiplexed genome editing. We demonstrate
( u6 F6 o- H* m. O7 Y& U+ Rthat CRISPR/Cas-mediated gene editing allows the2 d/ i- P* Q" M$ `
simultaneous disruption of five genes (Tet1, 2, 3,: o- ^8 v$ |- k2 [
Sry, Uty - 8 alleles) in mouse embryonic stem (ES)
% L9 i+ ]& }& ccells with high efficiency. Coinjection of Cas9
' q- W3 N3 Z8 w! UmRNA and single-guide RNAs (sgRNAs) targeting u$ U1 u% x& G+ s+ ]
Tet1 and Tet2 into zygotes generated mice with biallelic: t. t }. O7 Y4 I
mutations in both genes with an efficiency of
$ w8 `: f4 M1 n% n* e: i8 F1 B/ v80%. Finally, we show that coinjection of Cas9
8 q! T7 {0 H$ i0 {( \% C! ]: B( WmRNA/sgRNAs with mutant oligos generated precise
* S6 i( ]3 u# c1 {9 O% t& Ipoint mutations simultaneously in two target
4 z: n6 |, g; y: E% N9 {genes. Thus, the CRISPR/Cas system allows the
8 q+ K& B& q6 qone-step generation of animals carrying mutations
: w0 Q, T5 ~( z8 D# Y0 B9 Zin multiple genes, an approach that will greatly accelerate; o6 ~3 q" q6 d% t6 ^9 j: m! V3 j9 Y% `
the in vivo study of functionally redundant7 w H, E3 X( m* B7 }) N% m
genes and of epistatic gene interactions. |
-
总评分: 威望 + 5
包包 + 20
查看全部评分
|
发表于 2013-5-9 23:38
发表于 2013-5-10 10:26
