屬于植物DNA內(nèi)切核酸酶CEL

2015-11-14IMPROVING SPECIFICITY

OFCRISPR/CAS9OUTLINESurveyor

nuclease

assayKnockin

strategy

via

Crispr/Cas9Specificity

of

Crispr/Cas9TALEN

andCRISPR/CAS9Strategies

for

improving

Crispr/Cas9Evolution

of

Crispr/Cas9SURVEYOR

NUCLEASESurveyor

nucleaseSURVEYOR

NUCLEASECruiserTMKNOCKIN

STRATEGY

VIA

CRISPR/CAS9KNOCKIN

STRATEGY

VIA

CRISPR/CAS9KNOCKIN

STRATEGY

VIA

CRISPR/CAS9KNOCKIN

STRATEGY

VIA

CRISPR/CAS9在設(shè)計(jì)HR模板時(shí)可以參考下面的建議：?Cell

paper:/S0092-8674(13)01015-5h

/?KNOCKIN

STRATEGY

VIA

CRISPR/CAS9在設(shè)計(jì)HR模板時(shí)可以參考下面的建議：??Cell paper:

h

//S0092-8674(13)01016-7?SPECIFICITY

OF

CAS9方法SPECIFICITY

OF

CAS9SPECIFICITY

OF

CAS9SPECIFICITY

OF

CAS9??SPECIFICITY

OF

CAS9??SPECIFICITY

OF

CAS9SPECIFICITY

OF

CAS9SUMMARYSPECIFICITY

OF

CAS9STRATEGIES

FOR

IMPROVINGCRISPR/CAS9?STRATEGIES

FOR

IMPROVINGCRISPR/CAS9?STRATEGIES

FOR

IMPROVINGCRISPR/CAS9?STRATEGIES

FOR

IMPROVINGCRISPR/CAS9?STRATEGIES

FOR

IMPROVING

CRISPR/CAS9?STRATEGIES

FOR

IMPROVING

CRISPR/CAS9?STRATEGIES

FOR

IMPROVING

CRISPR/CAS9?SUMMARY?TALEN

VS

CRISPR-CAS9識(shí)別原理TALEN根據(jù)一個(gè)重復(fù)單元對(duì)應(yīng)一個(gè)堿基，將多個(gè)（16）重復(fù)單元串聯(lián)來(lái)決定識(shí)別序列的特異性。CRISPR/Cas9依賴于crRNA或sgRNA上的20個(gè)堿基的向?qū)蛄信c目的DNA，以及PAM序列決定切割位點(diǎn)的特異性。利用CRISPR/Cas系統(tǒng)可高效切割，還可以同時(shí)對(duì)多個(gè)

組的位點(diǎn)進(jìn)行切割和修飾。識(shí)別原理TALENCRISPR/Cas9質(zhì)粒構(gòu)建T

C

A

T

C

A

G

C

T

C

A

T

G

C

A

G

G

CTALEN比CRISPR/Cas9切割位點(diǎn)的特異性高。切割點(diǎn)的特異性TALEN切割位點(diǎn)的特異性由兩個(gè)TALEN識(shí)別序列（各16個(gè)單元）共同決定。CRISPR/Cas9系統(tǒng)切割位點(diǎn)的特異性取決于向?qū)蛄校?0bp）和PAM（NGG）序列。EVOLUTION

OF

CRISPR/CAS9Fusion

of

catalytically

inactive

Cas9

to

FokI

nucleaseimproves

the

specificity

of

genome

modificationEVOLUTION

OF

CRISPR/CAS9EVOLUTION

OF

CRISPR/CAS9EVOLUTION

OF

CRISPR/CAS9EVOLUTION

OF

CRISPR/CAS9A

split-Cas9

architecture

for

inducible

genome

editingand

transcription

modulationEVOLUTION

OF

CRISPR/CAS9A

Light-inducible,

RNA-guided

activation

ofendogenous

human

genes

by

the

LACE

systemCRISPR/CAS9的應(yīng)用：敲除小鼠CRISPR/CAS9的應(yīng)用：敲除小鼠???CRISPR/CAS9的應(yīng)用：小鼠肝臟突變誘導(dǎo)CRISPR/CAS9的應(yīng)用：小鼠肝臟突變誘導(dǎo)CRISPR/CAS9的應(yīng)用：小鼠肝臟突變誘導(dǎo)CRISPR/CAS9的應(yīng)用：哺乳動(dòng)物腦敲除CRISPR/CAS9的應(yīng)用：哺乳動(dòng)物腦敲除CRISPR/CAS9的應(yīng)用：哺乳動(dòng)物腦 敲除CRISPR/CA