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本帖最后由 zwb96 于 2013-5-9 23:41 编辑 0 g. y3 e' `7 T$ N) k
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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。. r" z2 F2 R) B1 a; N+ G6 S0 [ n
Mice carrying mutations in multiple genes are traditionally2 p, P" ^2 m4 v
generated by sequential recombination in+ |3 f$ P( U& N* l! {/ j- W
embryonic stem cells and/or time-consuming intercrossing: _6 T" F& L- r& ]/ E( W
of mice with a single mutation. The' g6 {( C' z, D& N
CRISPR/Cas system has been adapted as an efficient
- d9 u+ {# n) Lgene-targeting technology with the potential
5 e) ^ {# ]1 | `, t' Ifor multiplexed genome editing. We demonstrate5 U2 [* v; [. l
that CRISPR/Cas-mediated gene editing allows the
5 C. T5 m, F! p4 O+ Qsimultaneous disruption of five genes (Tet1, 2, 3,5 l. _. N( R- \1 l$ \( H
Sry, Uty - 8 alleles) in mouse embryonic stem (ES)
/ U% o6 W4 |1 I) M$ E/ Ocells with high efficiency. Coinjection of Cas9
# a0 _6 @" G0 fmRNA and single-guide RNAs (sgRNAs) targeting
- ?8 C- s! F c* N5 NTet1 and Tet2 into zygotes generated mice with biallelic
: K, Q' q1 K+ G, vmutations in both genes with an efficiency of1 _3 [% j% _9 B5 a/ W1 T
80%. Finally, we show that coinjection of Cas9
1 {1 t% {7 J9 S0 jmRNA/sgRNAs with mutant oligos generated precise
8 I' y, a8 Z [% s1 e% v: kpoint mutations simultaneously in two target
5 L% L m+ R# j) w. [genes. Thus, the CRISPR/Cas system allows the
& M$ A/ r8 p/ _+ _- P }. s( tone-step generation of animals carrying mutations% X. r4 ]. H6 S# q' ~& k& t
in multiple genes, an approach that will greatly accelerate+ P9 g, F: [6 L. n
the in vivo study of functionally redundant' a( [" v4 Z8 i! ~
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
