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INTRODUCTION
11
willcertainlybeheraldedasoneofthemostimportantscientifcendeavors
ofourtimeż
Quiterecently,anew,powerfulmethodofgeneengineeringwasde-
velopedżThemethod,namedCRISPR-Cas(Figż1),isanefcientandreliable
waytomakeprecise,targetedchangestothegenomeoflivingcellsż
CRISPR(clusteredregularlyinterspacedshortpalindromicrepeats)and
CRISPR-associated(Cas)genecodingfornucleaseareessentialinadaptive
immunityofbacteriaandarchaea,enablingtheorganismstorespondto
1
3
guideRNA
targetDNA
Cas9
guideRNA
THECRISPR/Cas9TECHNIQUE
EDITINGAGENEUSING
2
4
Cas9
editedDNA
guideRNA
Fig.1.AnoverviewoftheCRISPR-Cas9method(CRISPRgeneeditingmethod)ż
1)AguideRNAwithafragmentmatchingthetargetDNAsequence(red)isdesigned
andcreatedż2)TheguideRNAisaddedtothetargetcellalongwiththeCas9proteinż
3)GuideRNApairswiththematchingfragmentoftargethostDNA(green),whichgets
cutbytheCas9protein,creatingadouble-strandbreakataprecisetargetlocationż
4)DesiredDNAsequencemodifcationscanbeintroducedatthepreciselocation
withinthegenomeż