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本帖最后由 zwb96 于 2013-5-9 23:41 编辑
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7 L8 n, u E+ J% _; ?: |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。
7 }$ y3 O5 w8 {* K. h! TMice carrying mutations in multiple genes are traditionally
' i: n9 G. } u& bgenerated by sequential recombination in
/ ~6 Q/ h( F* ^' }3 Jembryonic stem cells and/or time-consuming intercrossing$ z1 ~2 N" V3 ?# {5 F2 t Z
of mice with a single mutation. The
; r$ Z- r- w: P6 v$ J" YCRISPR/Cas system has been adapted as an efficient8 s; S6 P# S9 Q1 ?
gene-targeting technology with the potential/ }6 x/ m5 h: P9 s6 s) d$ A
for multiplexed genome editing. We demonstrate# K) w4 y. ~' ^7 o0 m; l6 A
that CRISPR/Cas-mediated gene editing allows the
) U9 O6 a3 g- f, s! k0 Z6 S# E. b9 Ssimultaneous disruption of five genes (Tet1, 2, 3,
5 `- n: x: h; E: m7 y8 gSry, Uty - 8 alleles) in mouse embryonic stem (ES)
) F; ]8 W9 X/ {' Q+ p1 Rcells with high efficiency. Coinjection of Cas9* O+ B- K4 M/ C1 |& q
mRNA and single-guide RNAs (sgRNAs) targeting) I( O: Z: F1 g9 t
Tet1 and Tet2 into zygotes generated mice with biallelic B. J. B+ m/ {$ U% a1 Q( c
mutations in both genes with an efficiency of
; v+ O$ R6 R- F, \& _: F80%. Finally, we show that coinjection of Cas9 x) u, F+ H$ }: @! }) H" T0 ]- X: h
mRNA/sgRNAs with mutant oligos generated precise
& I2 u+ ^# Q7 l/ Epoint mutations simultaneously in two target
$ J; W# |" E* s8 Ogenes. Thus, the CRISPR/Cas system allows the, M5 }. P" P) C9 t+ C1 e' h6 m
one-step generation of animals carrying mutations6 P3 h/ ^- u+ ^+ {1 \/ m$ z" g# \8 o
in multiple genes, an approach that will greatly accelerate
3 @& z1 d6 p" o0 m7 J; I w' Othe in vivo study of functionally redundant: {( A* a1 D! V. b$ w; o* p
genes and of epistatic gene interactions. |
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发表于 2013-5-9 23:38
发表于 2013-5-10 10:26
