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本帖最后由 zwb96 于 2013-5-9 23:41 编辑
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) X. b; j8 O2 r! j( d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。
9 f) }, _/ T! [- f; ~- V4 l$ sMice carrying mutations in multiple genes are traditionally4 C: y+ m& Y! I8 V- Y2 s6 X7 g
generated by sequential recombination in1 r4 P5 D* U' V3 {
embryonic stem cells and/or time-consuming intercrossing4 u [2 d6 Q; w+ g" J
of mice with a single mutation. The1 J+ k! S: O6 b
CRISPR/Cas system has been adapted as an efficient0 U: S( ~! F4 |( C5 Y, X) J4 j
gene-targeting technology with the potential+ V: C6 Q% t( @
for multiplexed genome editing. We demonstrate
2 r S( M8 D% D# Kthat CRISPR/Cas-mediated gene editing allows the
p0 V4 k! [1 ? Dsimultaneous disruption of five genes (Tet1, 2, 3,0 O! J; [0 Y2 V0 W. u2 [& v
Sry, Uty - 8 alleles) in mouse embryonic stem (ES)
+ I: s+ q( k# g6 d3 q* a' mcells with high efficiency. Coinjection of Cas9
3 G- O- _, q4 t/ y6 j* UmRNA and single-guide RNAs (sgRNAs) targeting7 U0 v% Y: K% b& z9 L( [
Tet1 and Tet2 into zygotes generated mice with biallelic
6 n' t* @ ^6 e4 m) ~) Nmutations in both genes with an efficiency of& S% A. s- D8 q9 J7 S& [9 }8 w( J
80%. Finally, we show that coinjection of Cas95 V% O1 A6 f6 N3 r; ]* U1 ]
mRNA/sgRNAs with mutant oligos generated precise, ~4 a1 I- ~3 d2 j( |* Y
point mutations simultaneously in two target
; b& k3 D" @9 w: jgenes. Thus, the CRISPR/Cas system allows the
4 d, X2 m0 o; r" k& |# Done-step generation of animals carrying mutations: B, N& o3 `- r# o' z5 C
in multiple genes, an approach that will greatly accelerate3 I" w% h6 s/ w
the in vivo study of functionally redundant
" K8 j5 K( K: `+ z9 Jgenes 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
