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7/25/2019 Chapter 31 Regulatory Rna
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CHAPTER 31 REGULATORY RNA
30.4 BACTERIA CONTAIN REGULATOR RNA
1. Small untranslate ant!"sense RNA s#e$!es $an re%ulate
%ene eress!'n !n ('t) (a$ter!a * mammal!an $ells.
+. C'ntr'l '$$urs !n man, le-els trans$r!#t!'n term!nat!'n/
RNA #r'$ess!n% * e%raat!'n/ translat!'n !n!t!at!'n.
3. sRNA 'ten en$'e at t)e same l'$! as tar%et RNA
re%ulate (, (ase #a!r!n% at t)e re%!'ns '
$'m#lementar!t,.4. Ba$ter!al re%ulat'r RNAs are $alle sRNAs.
2. Se-eral sRNAs are ('un (, #r'te!n Hfq)!$) !n$reases
t)e!r ee$t!-eness.. Tandem repeats$an (e trans$r!(e !nt' #'erul
ant!-!ral RNAs 5!mmun!t,6
7. O&,S sRNA !nE.colia6 O&,S 8 a(unant/ 109 nt/ untranslate RNA !nu$e
(, '&!at!-e stress
(6 O&,S as #le!'tr'#!$ re%ulat'r re%ulate eress!'n '
mult!#le %enesa$t!-ates:re#ress eress!'n ' ;40
l'$! at post-transcriptional level
$6 O&,S as ant!"mutat'r/ #r'te$t $ells r'm s#'ntane'us
an $)em!$all, !nu$e muta%enes!s.
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1+. CRISPR nu$lease s,stem $'ns!sts ' 3 $'m#'nents Cas9/
$rRNA * tra"$rRNAa6 Gu!e RNA 5s%RNA6 8 $rRNA * tra"$rRNA
13. Cas9 8 CRISPR ass'$!ate #r'te!n $'nta!n + nu$lease
'ma!ns -ar, : small $rRNAsa6 In-'l-e !n se=uen$e re$'%n!t!'n * s"NA $lea-a%e
(6 PAD 8 #r't's#a$er"aa$ent m't! 5NGG6
14. Inu$e 'u(le stran (rea leas t' NA re#a!r -!a
a6 Homolo"$ directed repair %H&R'!n t)e #resen$e ''n'r tem#late : )'m'l'%, t' t)e tar%ete l'$us
#re$!se %ene e!t!n%:re#a!r
(6 Non-homolo"ous end (oinin" %NH)*'/ a(sen$e '
'n'r tem#late!nsert!'ns:elet!'ns 5!nels6/
!sru#t tar%et l'$us
30.2 C.ELEGANS AS DOEL SYSTED
1. ?eatures ' C. elegansa6 Eas!l, $ult!-ate 5%r' 'n #etr! !s) ee 'n n'n"
)a@ar'us (a$ter!a/ $)ea# t' ma!nta!n6
(6 S)'rt %enerat!'n t!me 3 a,s r'm e%% t' ault)''$6 Small 1mm
6 ?e:!&e n'. ' $ells emale 5/ 9296/ male 5O/
10316
e6 Amena(le 'r %enet!$ anal,s!s" E!$!ent re#r'u$t!-e s,stem Da!nta!ne as
)erma#)r'!tes 5re#r'u$t!-e 'r%ans ass'$!ate
: ('t) male * emale6" Dale e&!st 'r %enet!$ stu!es
" Small %en'me s!@e 100D(
6 Trans#aren$, e-el'#ment anal,s!s r'm a s!n%le
$ells an all $ell t' (e l!nea%e
+. Clea-a%e e-ents * $ell l!nea%e e-el'#ment
3. !s$'-er!es ' %enet!$ re%ulat!'n ' 'r%an e-el'#ment *
#r'%ramme $ell eat) 5t' ma!nta!n !&e n'. ' $ells6
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30. RNA INTER?ERENCE BY SYNTHETIC SDALL
INTER?ERING SRNA 5SIRNA6
1. !s$'-er, ' small !nterer!n% RNA 5s!RNA6 !n C.
elegans/ res#'ns!(le 'r RNA !ntereren$e 58 %ene
s!len$!n% (, dou!le-stranded RNA6
+. Eer!ment !n!n%s
a6 Ine$t!'n ' ant!"sense RNA t' s!len$e a %ene !n C.
eleganst' assess %ene un$t!'n'r
(6 Ine$t!'n ' sRNAm're #'tent ee$t
3. !$er 8 an en'nu$lease t)at #r'$ess 'u(le"strane
#re$urs'r RNA t' +1"+3 nu$le't!es RNA !ntereren$e
m'le$ules
?un$t!'nal 'ma!ns !n !$er !n$lue )el!$ase/ U?+
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2. A##l!$at!'n ' s!RNA !n (!'me!$al resear$) Useul *
ee$t!-e t''l 'r s#e$!!$ %ene n'$'n. L!m!tat!'ns '"tar%et ee$t
7. D!n!m!@at!'n ' '"tar%et ee$ts
a6 Careul es!%n ' s)RNA $'nstru$t
(6 Intr'u$e + 'r m're s!RNA$6 Res$ue unes!re #)en't,#e $ause (, an s!RNA :
an s!RNA res!stant eress!'n $'nstru$t
30.7 DICRO"RNA ARE IESPREA REGULATORS IN
EUARYOTES
1. D!$r'"RNA 5m!RNA6 are en'%en'us ;++nt RNA
re%ulate %ene eress!'n !n #lant/ an!mal * un%!5euar,'tes6
+. m!RNA 'r!%!nate r'm #r!"m!RNA 8 2F"$a##e * 3F"
#'l,A ull len%t) #re$urs'rs s,nt)es!@e (, RNA #'l II.
3. m!RNA !t)!n a %en'm!$ $luster are 'ten relate t' ea$)
't)er.
4. One m!RNA $an !n)!(!t '-er 100 !erent tar%et %enes.2. !erent t,#es ' m!RNA
a6 stRNA5s)'rt tem#'ral RNA6 m'ulates mRNA
eress!'n ur!n% e-el'#ment(6 piRNA re%ulate %ene eress!'n !n %erm $ells an
a$t t' s!len$e trans#'sa(le elements
$6 siRNA $'m#lementar, t' -!rus * trans#'sa(le
elements
. ?!rst m!RNA lin-+, let-5stRNA6 ere !ent!!e !n
C.elegans 5?'un!n% mem(ers ' an a(unant $lass '
t!n, RNAs 'rl6
7. P'st"trans$r!#t!'nal re%ulat!'n ' )eter'$)r'n!$ %ene lin-
14(, lin-4me!ates tem#'ral #attern 'rmat!'n !n
C.elegansa6 Ea$) sta%e ' e-el'#ment 5$,$le ' $ell !-!s!'ns/
%r't) ' t!ssue/ 'rmat!'n ' 'r%ans6 re=u!re #r'#er
t!m!n%(6 Heter'$)r'n, 8 $)an%e !n t)e relat!-e t!m!n% '
e-el'#mental e-ents.
$6 + %eneral #)en't,#es seen !n )eter'$)r'n!$ mutants
" Precocious e-el'#mental e-ents are s!##e
" Retarded e-el'#mental e-ents are re#eate6 Heter'$)r'n!$ mutat!'n
" Ae$t !erent t!ssues !ntest!ne/ e#!erm!s/
mus$le/ neur'ns
" Ae$t !erent e-el'#mental e-ents $ell
!-!s!'n #attern/ $ell $,$le len%t)/ !erent!at!'n
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lin-4, let-7 RNA #re$urs'r 'rm a )a!r#!n stru$ture mature
m!RNA !s s)'n as (lue 5lin-46 * re 5let-76
1+. A$t!'n ' lin-4 & let-7
a6 Base #a!r!n% ' m!RNAs 52F"3F6 t' t)e / 0TR'
tar%et mRNAs 5lin-14, lin-416(6 B!n!n% s!te (eteen lin-41 & let-7 ma,(e
re$'%n!@e (, RBP 5RNA (!n!n% #r'te!ns6
13. let-7$'ntr'ls L4"t'"ault trans!t!'n !n C. elegans. lin-14
!s 'ne tar%et. let-7 mutant seam $ells a!l t' term!nall,
!erent!ate an $'nt!nue t' !-!e(urst!n% t)r'u%)
-ul-a
14. let-7 !s $'nser-e a$r'ss se-eral s#e$!es m!RNAre%ulat!'n ' %ene eress!'n !s n't e&$lus!-e t'
C.elegans
m!RNA !s )!%)er euar,'tes
30.< PROCESSING O? DIRNA IN EUARYOTES
1. Small RNAs are #r'u$e (, #r'$ess!n% ' l'n%er
#re$urs'rs. 5#r!mar,6 #r!"m!RNA#re"m!RNA
m!RNA u#le&
+. &rosha 1 dicer are RNase III 5en'nu$lease6 ('t)
#r'$ess 'u(le"strane l'n% RNA trans$r!#ts3. &rosha $'n-ert #r!"m!RNA t' #re"m!RNA
a6 O$$urs !n nu$leus(6 Clea-a%e s!tes are 11(# aa, r'm sRNA * ssRNA
un$t!'ns lea-e a +"nu$le't!e '-er)an% at 3F en
$6 Pre"m!RNA ater r's)a $lea-a%e !s 2"70nt l'n%
4. &icer $'n-ert #re"m!RNA t' mature m!RNA
a6 O$$ur !n $,t'#lasm mature m!RNA !s %enerate !n
$,t'#lasm
(6 Clea-a%e als' lea-es +"nu$le't!e '-er)an% at 3F en
$6 m!RNA ater !$er $lea-a%e !s +1"+3nt l'n%
2. RISC5RNA"!nu$e s!len$!n% $'m#le&6 8 a
r!('nu$le'#r'te!n #art!$le $'m#'se ' a s)'rt s!n%le"
strane s!RNA an a nu$lease t)at $lea-es mRNAs
$'m#lementar, t' s!RNA 5re$e!-e s!RNA r'm !$er an
el!-ers !t t' mRNA6
. C'm#'nents !n RISC
a6 Ar"onauts protein54 !n mammals/ all !n-'l-e !n
m!RNA me!ate translat!'n re#ress!'n 'nl, A%'"+
!n-'l-e !n mRNA $lea-a%e6
(6 2uide RNAa stran !n mature m!RNA t)at !s
!n$'r#'rate !nt' RISC 2Fen : less (# !ll (e
$)'sen #reerent!all,
$6 Passen"er RNA 't)er stran ' mature m!RNA t)at
!s e%rae7. Generat!'n ' RISC
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30.< RNA INTER?ERENCE DOE O? ACTION
1. m!RNA re%ulate %ene eress!'n -!a (ase #a!r!n% :$'m#lementar, se=uen$es !n tar%et mRNA
+. RNA !ntereren$e tr!%%er e%raat!'n/ translat!'n
!n)!(!t!'n 'r mRNA a$t!-at!'n.3. m!RNA m'e ' a$t!'ns !n !erent s#e$!es
a6 Plants mRNA e%raat!'n
(6 An!mals translat!'nal re%ulat!'n
$6 Plant * an!mals/ ,east trans$r!#t!'nal re%ulat!'n
5)!st'ne met),lat!'n t' !na$t!-ate $)r'mat!n6
4. De$)an!sms ' m!RNA me!ate %ene s!len$!n%
a6 In)!(!t!'n ' translat!'n el'n%at!'n
(6 C'"translat!'nal #r'te!n e%raat!'n$6 C'm#et!t!'n 'r t)e $a# stru$ture
6 In)!(!t!'n ' r!('s'mal su(un!t '!n!n%
e6 In)!(!t!'n ' mRNA $!r$ular!@at!'n -!a eaen,lat!'n
6 Pr'm'te eaen,lat!'n * e$a##!n%
2. 'u(le"strane RNA 5!ne$te r'm e&ternal s'ur$e6
!n)!(!t translat!'n/ tr!%%er mRNA e%raat!'n !n
mammal!an $ells 5se=uen$e s#e$!!$ ee$ts6 !t $an als'
s!len$e )'st %enes
30.9 HETEROCHRODATIN ?ORDATION REJUIRES
DIRNA
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