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T*
[(ge)16.7(n)12.4(i)0(t)19.6(o)0(r)-597.6(c)0(e)20.3(l)0(l)17(s)-604.2(\(H)17(S)12.6(P)10.2(Cs)16.2(\))]TJ
/F7 1 Tf
10.9585 0 TD
[(ex)-592.2(vi)18.4(vo)]TJ
/F5 1 Tf
3.9888 0 TD
[(to)-600(t)11(r)0(e)20.3(a)0(t)-596.7(X)0(-)22.1(l)0(i)17(n)0(k)20.1(e)0(d)]TJ
-14.9473 -1.1869 TD
[(ad)18.4(re)20.3(no)19.2(le)20.7(uk)14.7(o)12.2(d)0(y)18.2(s)0(t)19.2(o)12.2(ph)15.4(y,)-343.2(me)19.3(t)11(a)0(c)18.1(h)0(r)15(o)12.2(m)12.4(a)0(t)15.7(i)0(c)-346.6(l)0(e)20.7(u)0(k)20.1(o)0(d)15.2(y)0(s)23.5(t)0(r)19.1(o)0(-)]TJ
0 -1.1923 TD
[(ph)20.8(y,)-614.2(an)17(d)-620.3(W)0(i)17.4(s)13.6(ko)19.9(tt)16.7(A)18.1(ld)22.3(ri)16.5(ch)-613.7(sy)18.1(n)12.4(d)0(r)16.4(o)12.2(me)19.3(.)]TJ
7.8454 0 0 6.9729 247.4645 157.096 Tm
[(22)27.3(2)20.1(4)]TJ
10.4607 0 0 10.4607 275.244 152.4472 Tm
[(L)10.2(e)12.2(nt)18.1(i-)]TJ
-20.3832 -1.1923 TD
[(vi)18.4(ru)20.4(s)-317(v)0(e)22.1(c)0(t)19.1(o)0(r)20.3(s)-317(ar)18.1(e)-312.9(a)0(l)24(s)0(o)-304.8(b)0(e)16.9(i)13.9(ng)-308.3(s)13.6(u)0(c)15(c)0(e)20.3(s)0(s)21.8(f)0(u)14.8(l)13.9(ly)-306.8(ap)18.4(pl)22.3(ie)15.3(d)-311.4(i)0(n)]TJ
0 -1.1869 TD
.3594 Tc
[(t)370.4(h)359.4(ec)359.4(l)375.9(i)359.4(n)380.3(i)359.4(cf)359.4(o)374(r)-6.5(t)370.4(h)359.4(ep)359.4(r)375.8(o)371.6(d)359.4(u)374.8(c)372.8(t)359.4(i)379(o)359.4(no)359.4(fc)359.4(a)377.5(n)371.8(c)359.4(e)374.2(r)372.8(-)359.4(s)375.4(p)359.4(e)380(c)359.4(i)375.9()371.8(c)-12(c)359.4(h)379.8(i)359.4(-)]TJ
0 -1.1923 TD
0 Tc
[(me)19.3(ri)16.5(c)-203.4(a)0(n)17.1(t)11(ig)18.4(e)12.2(n)-204.4(re)14.8(c)13.4(e)0(p)15.2(t)11(o)12.2(r)0(-)15.8(e)12.2(xp)18.2(re)20.3(ss)16.3(in)20.9(g)-206.9(T)-206.6(ly)18.4(m)12.4(p)0(h)15.4(o)12.2(cy)17.9(t)11(e)0(s)-196.4(t)11(o)]TJ
T*
[(t)11(r)0(e)14.8(a)0(t)-331.2(l)0(e)20.7(u)0(k)14.7(e)12.2(mi)21(a)]TJ
7.8454 0 0 6.9729 132.7748 107.2629 Tm
(25)Tj
10.4607 0 0 10.4607 144.9637 102.6141 Tm
[(an)22.5(d)-349.3(g)0(l)18.4(i)13.9(ob)16.9(la)18.5(s)13.6(t)0(o)17.9(m)0(a)17.1(.)]TJ
7.8454 0 0 6.9729 229.4929 107.2629 Tm
-.0173 Tc
(26)Tj
10.4607 0 0 10.4607 241.6251 102.6141 Tm
0 Tc
[(F)10.2(e)0(a)16.9(s)13.6(ib)18.5(il)17(it)19.6(y)-342.4(o)0(f)]TJ
-17.1693 -1.1869 TD
[(t)11(h)0(e)-490.2(l)0(e)20.7(n)0(t)18.1(i)0(v)18.4(i)0(r)16.5(a)0(l)-485.5(a)0(p)18.4(p)0(r)16.4(o)12.2(ac)18.1(h)-497(f)0(o)20.1(r)-501.4(C)13.5(R)0(I)18.4(S)0(P)17.4(R)13.5(/C)18.1(as)18.2(9)-499.4(i)13.9(s)-501.3(s)0(u)20.6(p)0(-)]TJ
0 -1.1923 TD
[(po)20.6(rt)19.1(ed)-196.2(by)-196.9(st)19.2(ud)20.8(ie)15.3(s)-203.2(d)0(e)20.6(m)0(o)19.3(n)0(s)20.6(t)0(r)19.1(a)0(t)15.7(i)0(n)20.9(g)-206.9(su)20.6(cc)16(e)12.2(s)0(s)16.3(f)13.2(ul)-195.9(de)15.2(l)13.9(i)0(v)18.4(e)0(r)20.3(y)]TJ
T*
[(t)11(o)-388.8(m)12.4(a)0(m)17.1(m)12.4(al)18.5(ia)18.5(n)-383.2(c)0(e)20.3(l)0(l)17(s)-387.5(bo)22.3(t)11(h)]TJ
/F7 1 Tf
11.9015 0 TD
[(ex)-375.4(v)10.6(i)0(v)18.4(o)]TJ
/F5 1 Tf
3.5553 0 TD
[(an)22.5(d)]TJ
/F7 1 Tf
2.1028 0 TD
-.0119 Tc
[(in)-400.5(vi)-11.9(v)6.5(o)]TJ
/F5 1 Tf
3.1596 0 TD
.3856 Tc
[(.T)395.8(h)398(e)]TJ
-20.7192 -1.1869 TD
0 Tc
[(di)16.9(ve)22.1(rs)16.2(e)-356.3(a)0(p)18.4(p)0(l)16.9(i)13.9(ca)18.1(ti)19.6(on)19.2(s)-360.4(o)12.2(f)-360.7(l)0(e)20.7(n)0(t)18.1(i)0(v)18.4(i)0(r)16.5(u)12.4(s)-360.4(v)0(e)22.1(c)0(t)19.1(o)0(r)20.3(s)-360.4(fo)20.1(r)-360.5(d)0(e)20.6(l)0(i)17(v)0(-)]TJ
0 -1.1923 TD
[(e)12.2(r)0(y)-534.9(o)12.2(f)-545(C)0(a)18.1(s)13.6(9/)20.1(gR)17.9(NA)-534.9(c)13.4(o)0(m)19.3(p)0(o)15.2(n)12.4(e)12.2(n)0(t)18.1(s)-544.6(in)20.9(cl)16.5(u)12.4(d)0(e)-532.2(m)0(o)19.3(d)0(e)20.6(l)0(i)17(n)12.4(g)]TJ
25.4288 64.2387 TD
[(ca)18.1(n)12.4(c)0(e)14.8(r)-284.6(in)-271.7(mi)15.5(c)13.4(e)0(,)]TJ
7.8454 0 0 6.9729 399.4015 716.9951 Tm
-.018 Tc
[(27)-18(30)]TJ
10.4607 0 0 10.4607 423.6093 712.3463 Tm
0 Tc
[(de)20.6(ve)16.7(lo)20.7(pm)20.8(en)19.2(t)-287(o)12.2(f)-284.8(s)0(e)20.4(l)0(e)20.7(c)0(t)13.6(i)13.9(on)-278.8(ba)20.1(s)13.6(e)0(d)]TJ
-9.1375 -1.1923 TD
[(ge)16.7(n)12.4(e)-220.8(di)16.9(s)13.6(r)0(u)15(p)13.8(ti)14.1(o)12.2(n)-220.6(as)18.2(s)13.6(a)0(y)19.9(s)-224.9(t)11(o)-220.8(c)13.4(o)0(n)19.2(d)0(u)15.4(c)13.4(t)-222(c)0(o)20.3(m)0(p)15.4(l)13.9(ex)-210.9(ga)19.9(in)-212.1(an)17(d)]TJ
0 -1.1869 TD
[(lo)20.7(ss)-422.6(o)12.2(f)-431.2(f)13.2(u)0(n)19.4(c)0(t)19.1(i)0(o)15.3(n)-426.6(ge)16.7(n)12.4(e)12.2(ti)14.1(c)-425.6(s)0(c)16.2(r)0(e)20.3(e)12.2(ns)15.2(,)]TJ
7.8454 0 0 6.9729 483.477 692.1069 Tm
-.0261 Tc
[(1)-8.8(3)-26.1(,3)-26.1(1)-26.1(33)]TJ
10.4607 0 0 10.4607 519.7605 687.4581 Tm
0 Tc
[(an)22.5(d)-430.6(e)12.2(ra)18.1(di)16.9(-)]TJ
-18.3291 -1.1923 TD
[(ca)18.1(t)11(i)0(o)20.7(n)-280.2(of)-267.2(la)18.5(te)17.9(nt)-269.2(vi)18.4(ra)18.1(l)-278.7(i)0(n)20.9(f)0(e)20.1(c)0(t)19.1(i)0(o)15.3(n)12.4(s)-279.1(s)0(u)20.6(c)0(h)-272.2(a)0(s)-269(H)0(I)19.6(V)0(,)]TJ
7.8454 0 0 6.9729 537.8455 679.6345 Tm
-.0173 Tc
(34)Tj
10.4607 0 0 10.4607 549.2975 674.9856 Tm
0 Tc
[(he)19.2(p-)]TJ
-21.1528 -1.1923 TD
[(at)21.1(it)19.6(is)-232.6(B)-235.9(v)0(i)18.4(r)0(u)15(s)13.6(,)]TJ
7.8454 0 0 6.9729 392.3148 667.162 Tm
[(35)20.1()17.3(3)0(7)]TJ
10.4607 0 0 10.4607 416.0692 662.5132 Tm
[(an)17.1(d)-235.5(h)0(e)19.2(r)0(p)16.4(e)12.2(sv)18.1(ir)16.5(u)12.4(s)0(.)]TJ
7.8454 0 0 6.9729 495.4392 667.162 Tm
(38)Tj
10.4607 0 0 10.4607 506.4943 662.5132 Tm
[(In)-226.5(co)20.3(nt)18.1(ra)18.1(st)-230.1(t)11(o)]TJ
-17.0609 -1.1869 TD
[(t)11(h)0(e)-208.4(e)12.2(ff)15.7(e)12.2(c)0(t)19.1(i)0(v)18.4(e)-215.4(us)15.2(e)-215.4(o)12.2(f)-214.4(i)0(n)15.5(t)11(eg)16.7(r)13.4(a)0(t)15.7(i)0(n)20.9(g)-217.7(le)20.7(nt)18.1(iv)18.4(ir)16.5(u)12.4(s)-214(ve)16.7(ct)19.1(o)12.2(r)0(s)-211.4(f)13.2(or)]TJ
0 -1.1923 TD
[(ge)16.7(n)12.4(e)-258.7(re)20.3(pl)16.9(ac)18.1(e)12.2(m)12.4(en)19.2(t)-259.9(t)11(he)19.2(ra)18.1(py)18.2(,)-255.5(i)0(n)15.5(t)11(e)12.2(g)0(r)17.9(a)0(t)21.1(i)0(n)15.5(g)-255.7(ve)16.7(ct)19.1(o)12.2(r)0(s)-249.4(m)12.4(ay)]TJ
T*
[(no)19.2(t)-395.4(b)0(e)-378.8(s)0(u)15.2(i)0(t)19.6(a)0(b)20.1(l)0(e)-380.3(f)0(o)20.1(r)-398.5(de)20.6(li)17(ve)22.1(ry)-383.2(of)-386.4(Ca)18.1(s)13.6(9)0(,)-386.4(a)0(s)-382.8(t)11(he)-381.8(sa)18.2(fe)20.1(t)11.1(y)]TJ
0 -1.1869 TD
[(pr)16.4(o)12.2()0(l)15.5(e)-258.8(of)-256.3(s)13.6(u)0(s)15.2(t)11(ai)18.6(n)12.4(e)0(d)-255.8(C)13.5(as)18.2(9)-260.9(e)0(x)16.7(p)0(r)16.4(e)12.2(ss)21.8(io)15.3(n)-264(\()13.2(e.)16.8(g.)19.9(,)-261(o)0(f)20.1(f)0(-)15.6(t)11.1(ar)18.1(ge)22.1(t)]TJ
0 -1.1923 TD
[(cl)16.5(e)12.2(a)0(v)19.9(a)0(g)19.9(e)-231.7(an)22.5(d)-230.1(p)0(o)20.6(t)0(e)17.9(n)12.4(ti)14.1(al)-214.5(ef)14.6(f)13.2(e)0(c)20.2(t)0(s)-219.2(o)0(f)-218.4(C)0(a)18.1(s)0(9)-220.2(e)12.2(xp)18.2(re)20.3(ss)16.3(io)20.7(n)-231.5(o)12.2(n)]TJ
T*
[(pr)16.4(im)20.9(ar)18.1(y)-559.2(c)0(e)20.3(l)0(l)-552.1(h)12.4(om)19.3(eo)19.1(st)19.2(as)18.2(is)22.1(,)-559(f)0(u)14.8(n)12.4(ct)19.1(io)20.7(na)17.1(li)17(t)11(y)0(,)-549.2(a)0(n)17(d)-560.7(i)13.9(m)0(-)]TJ
0 -1.1869 TD
[(mu)19.5(ni)20.9(ty)15.5(\))-268.6(i)0(s)-259.7(n)0(o)19.2(t)-270.8(ye)16.7(t)-270.8(w)11.4(e)12.2(l)0(l)-264.8(u)12.4(nd)15.4(e)12.2(r)0(s)16.2(t)11(o)12.2(o)0(d)20.6(.)]TJ
1.1435 -1.1923 TD
[(Integrase-decient)-614.5(lentivirus)-621.1(vectors)-618.7(\(IDLVs\))]TJ
-1.1435 -1.1923 TD
[(are)-522.5(able)-517.4(to)-518.7(transduce)-521.6(primary)-520(human)-520.1(cells)-518.2(but)]TJ
0 -1.1869 TD
[(cannot)-454.1(integrate)-451.9(genetic)-451.3(cargo)-455.8(into)-454.4(the)-452(host)-449.3(cell)]TJ
0 -1.1923 TD
[(genome.)-200.3(Given)-205.7(the)-197.3(packaging)-207.8(capacity)-202.5(of)-202.1(lentivirus)]TJ
T*
[(vectors)-288.1(\()]TJ
/F12 1 Tf
3.8967 0 TD
(*)Tj
/F5 1 Tf
1.0026 0 TD
[(10)-164.2(kb\),)]TJ
7.8454 0 0 6.9729 410.9101 517.7195 Tm
(39)Tj
10.4607 0 0 10.4607 422.5889 513.0707 Tm
[(IDLVs)-287.8(are)-284(a)-288(useful)-285.2(tool)-286.6(for)-286.3(the)]TJ
-9.0399 -1.1869 TD
[(delivery)-458.9(of)-451.4(Cas9,)-457.7(gRNAs,)-455.4(and)-456.9(donor)-456.2(repair)-454.3(tem-)]TJ
0 -1.1923 TD
[(plates)-456.5(for)-459.7(homology-directed)-454.2(repair)-459.7(\(HDR\)-based)]TJ
T*
[(genome)-335(editing)-335.5(strategies.)-329.6(IDLVs)-336.6(have)-334.8(been)-332.4(used)]TJ
0 -1.1869 TD
[(to)-442.8(deliver)-436.3(zinc)-440.5(nger)-443.6(nucleases)-438.9(\(ZFNs\),)]TJ
7.8454 0 0 6.9729 524.5793 467.9432 Tm
(40,41)Tj
10.4607 0 0 10.4607 548.6739 463.2943 Tm
(TAL)Tj
-21.0931 -1.1923 TD
[(effector)-462.5(nucleases)-466(\(TALENs\),)]TJ
7.8454 0 0 6.9729 472.5353 455.4707 Tm
(42)Tj
10.4607 0 0 10.4607 486.0282 450.8219 Tm
(meganucleases,)Tj
7.8454 0 0 6.9729 561.4865 455.4707 Tm
(43)Tj
10.4607 0 0 10.4607 328.0251 438.3495 Tm
[(and)-364.8(donor)-358.6(repair)-362.1(templates)-361.1(for)-362.1(site-specic)-363.4(modi-)]TJ
0 -1.1869 TD
[(cation)-293.8(of)-288.9(primary)-292.4(target)-295.4(cells)]TJ
/F7 1 Tf
14.3132 0 TD
[(ex)-294.1(vivo)]TJ
/F5 1 Tf
3.4089 0 TD
(and)Tj
/F7 1 Tf
2.0269 0 TD
[(in)-294.1(vivo)]TJ
/F5 1 Tf
3.1109 0 TD
(.)Tj
-22.8599 -1.1923 TD
[(While)-355.9(the)-349.1(transient)-355.9(expression)-349.6(of)-353.9(IDLV-delivered)]TJ
T*
[(Cas9)-484(in)-488.5(rapidly)-485.7(dividing)-489.2(cells)-485.7(could)-487(represent)-487.1(a)]TJ
0 -1.1869 TD
[(safety)-260.5(advantage,)-259.1(in)-260.9(quiescent)-257.1(and)-261.8(slowly)-255.6(dividing)]TJ
0 -1.1923 TD
[(cells,)-524.4(including)-528.8(long-term)-528.6(HSPCs,)-521.6(neurons,)-529.9(and)]TJ
T*
[(hepatocytes,)-578.8(Cas9)-581.5(expression)-577.2(may)-580(persist)-580.4(after)]TJ
0 -1.1869 TD
[(IDLV)-268.9(delivery)-263.8(leading)-271.5(to)-264(undesired)-266.9(effects)-268.6(such)-267.9(as)]TJ
0 -1.1923 TD
[(higher)-277.9(off-target)-281.4(cleavage.)]TJ
/F2 1 Tf
9.9626 0 0 9.9626 328.0251 314.3621 Tm
[(AAV)-338.4(deli)-7.6(very)]TJ
/F5 1 Tf
10.4607 0 0 10.4607 339.9873 301.8897 Tm
[(Recombinant)-213.1(AAV)-219.8(\(rAAV\))-217.8(vectors)-212.2(are)-219(promising)]TJ
-1.1435 -1.1923 TD
[(gene)-527.7(delivery)-534.8(vectors)-526.5(for)-535.6(a)-526.5(spectrum)-531.7(of)-532.7(genetic)]TJ
0 -1.1869 TD
-.0001 Tc
[(disorders)-460(because)-452.6(of)-462.4(their)-457.3(nonpathogenicity,)-462.5(low)]TJ
0 -1.1923 TD
0 Tc
[(levels)-401.1(of)-381(im)15.5(m)12.4(u)0(n)19.4(e)-394.2(st)19.2(im)20.9(ul)20.9(at)15.7(io)20.7(n,)-384(ca)18.1(p)13.8(a)0(b)14.7(i)13.9(li)17(t)11(y)-396.6(t)11(o)-394.2(t)11.1(r)0(a)18.1(n)12.4(s-)]TJ
T*
[(du)20.8(ce)-640.9(bo)22.3(th)-637.7(di)16.9(vi)18.4(di)16.9(n)12.4(g)-645.9(an)17(d)-647.4(n)12.4(on)19.2(di)16.9(vi)18.4(di)16.9(n)12.4(g)-645.9(ce)14.8(ll)22.4(s,)-637.6(no)19.2(n-)]TJ
0 -1.1869 TD
[(in)20.9(te)17.9(gr)17.9(at)21.1(in)15.5(g)-629.6(n)12.4(at)21.1(ur)15(e)12.2(,)-629.5(an)17(d)-625.7(d)0(i)16.9(v)0(e)22.1(r)0(s)16.2(e)-627.3(t)11(i)0(s)16.7(s)0(u)20.6(e)0(-)20.1(t)0(a)15.7(r)13.4(ge)16.7(t)11(i)0(n)20.9(g)]TJ
0 -1.1923 TD
[(pr)16.4(o)12.2()0(l)15.5(e)12.2(s.)]TJ
7.8454 0 0 6.9729 365.7826 231.8173 Tm
-.0173 Tc
(44)Tj
10.4607 0 0 10.4607 376.6109 227.1684 Tm
0 Tc
[(AA)21.5(V)-219.6(i)0(s)-205.5(a)-217.6(no)19.2(ne)19.2(nv)16.9(e)12.2(l)0(o)15.3(p)13.8(ed)-212.4(p)13.8(a)0(r)18.1(v)0(o)16.7(v)0(i)18.4(r)13.4(us)-212.4(w)11.4(i)0(t)19.6(h)-220.6(a)]TJ
-4.6446 -1.1923 TD
[(pr)16.4(o)12.2(t)11(ei)15.3(n)-291.1(c)13.4(ap)18.4(si)16.7(d)-289.7(a)0(n)17(d)-289.7(re)20.3(la)18.5(t)11(i)0(v)18.4(e)12.2(ly)-285.1(s)13.6(m)0(a)17.1(l)0(l)-281.1(s)0(i)16.7(n)12.4(gl)18.4(e)12.2(-)0(s)16(t)11(ra)18.1(n)12.4(d)0(e)15.2(d)]TJ
0 -1.1869 TD
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[(delivery)-281.1(o)-2.4(f)-283.2(t)1.9(he)-284.2(CRISPR/)]TJ
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[(Cas9)-833.1(system,)-833.1(i)0(ncluding)-833.2(electropor)5(a)-13.9(t)12.6(ion,)-838.5(hydro-)]TJ
T*
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[(dynamic)-623.9(d)-.3(elivery,)-621.9(a)-14.1(n)13.8(d)-623.6(lipid-based)-623.6(nanoparticles)4.9(.)]TJ
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T*
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[(DN)-13.6(A)15.2(,)-556.4(R)5.3(N)-13.6(A)15.2(,)-556.4(o)4.1(r)-553(a)-550.9(c)-.2(om)4.3(plex)-551.1(of)-547.7(protein)-548.6(\()-.4(Cas9\))-553.2(a)1.9(nd)]TJ
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T*
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[(the)-186.7(C)-1.2(RISPR)-185.5(m)3.2(achinery)-189.1(f)4(o)-2.5(r)-185.5(t)1.8(ra)-14.7(n)13.2(s)-1.1(ient)-187.9(ex)-14.7(p)9(r)4.1(ession)-186.5(that)]TJ
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[(is)-267.1(suf)-2.7(cien)-2.7(t)-264.2(f)-1.9(or)-267.2(ge)-2.9(n)-2.7(e)-263(edit)-4.1(in)]TJ
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1.1435 -2.39 TD
0 Tc
(Electroporation.)Tj
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8.1619 0 TD
[(Originally)-554.7(described)-555.5(over)-553.8(30)]TJ
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[(years)-200.9(ago,)-196.3(the)-197.3(application)-197.4(of)-202.1(an)-199.7(electrical)-197.5(current)-198.8(to)]TJ
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[(cells)-193(in)-195.9(order)-195(to)-193.5(delivery)-193.3(DNA,)-193.7(or)-191.1(electroporation,)-192.5(is)]TJ
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[(now)-419.2(a)-423.5(commonly)-421.2(used)-419.5(research)-419.4(method.)-419.8(Electro-)]TJ
T*
[(poration)-366.9(effectively)-368.7(delivers)-368.5(DNA,)-367.1(RNA,)-365(and)-370.2(pro-)]TJ
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[(tein)-378.5(to)-372.3(cells)-377.3(and)-381.1(has)-375.8(been)-375.8(used)-376.1(more)-372.4(recently)-378.3(for)]TJ
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[(delivery)-290.9(of)-294.3(mRNA)-292.2(in)-293.4(a)-293.5(clinical)-290.6(setting,)-296(thus)-291.9(estab-)]TJ
T*
[(lishing)-316.4(the)-316.6(utility)-320.3(of)-316(this)-317.5(delivery)-318(method)-316(for)-318.8(clin-)]TJ
0 -1.1869 TD
[(ically)-353(based)-357.6(genome)-351.2(editing.)]TJ
7.8454 0 0 6.9729 446.2298 418.3936 Tm
(63)Tj
10.4607 0 0 10.4607 458.5889 413.7448 Tm
[(For)-354.3(delivery)-355.9(of)-353.9(the)]TJ
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[(CRISPR)-696(components,)-693.3(electroporation)-690.3(has)-695.6(been)]TJ
T*
[(utilized)-215.6(to)-209.8(deliver)-219.5(plasmid)-212(DNA)-214.5(encoding)-214(Cas9)-213(and)]TJ
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[(gRNAs)-313.7(to)-318.1(cancer)-315.2(cell)-315(lines,)-314.1(pluripotent)-315(stem)-313.8(cells,)]TJ
0 -1.1923 TD
[(and)-565.3(primary)-568.8(hematopoietic)-565.2(cells)-567(such)-565.9(as)-567.1(CD4)]TJ
/F11 1 Tf
7.8454 0 0 6.9729 545.329 368.5605 Tm
(+)Tj
/F5 1 Tf
10.4607 0 0 10.4607 307.8424 351.4393 Tm
[(T-cells)-353.8(and)-348.5(CD34)]TJ
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7.8454 0 0 6.9729 393.5621 356.0881 Tm
(+)Tj
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10.4607 0 0 10.4607 402.8597 351.4393 Tm
[(stem)-351.7(cells.)]TJ
7.8454 0 0 6.9729 453.9967 356.0881 Tm
(15,6467)Tj
10.4607 0 0 10.4607 490.1668 351.4393 Tm
(Electropora-)Tj
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[(tion)-221.3(has)-218.7(also)-221.2(been)-218.6(successfully)-223.6(used)-218.9(to)-220.6(deliver)-224.9(RNA)]TJ
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[(to)-421.1(fertilized)-422.1(mouse)-419.4(embryos)-420.1(in)-423.5(order)-422.6(to)-421.1(generate)]TJ
T*
[(genetically)-278.9(modied)-277.7(rodents.)]TJ
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(68,69)Tj
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10.4607 0 0 10.4607 319.8046 289.0771 Tm
[(Hydrodynamic)-244.4(delivery.)]TJ
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11.5004 0 TD
[(In)-199.4(preclinical)-196.9(models,)-199(a)]TJ
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[(method)-278.1(that)-275.2(has)-278.3(been)-278.2(successfully)-277.8(used)-278.5(for)]TJ
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[(in)-277.8(vivo)]TJ
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[(gene)-273(delivery)-274.6(is)-276(hydrodynamic)-275(delivery)-274.6(in)-277.2(which)-277.8(a)]TJ
T*
[(large)-309(volume)-306.2(of)-305.1(a)-309.7(DNA-containing)-307.1(solution)-304.6(is)-308.5(rap-)]TJ
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[(idly)-355(delivered)-356(via)-350.9(intravenous)-354.1(injection,)-350.6(resulting)]TJ
0 -1.1923 TD
[(in)-380.1(uptake)-377.2(of)-381(DNA)-377.1(by)-381.1(liver)-381(cells.)-378.1(Recently,)-379.1(hydro-)]TJ
T*
[(dynamic)-331.4(delivery)-334.3(was)-333.5(successfully)-332(used)-332.7(to)-334.4(deliver)]TJ
0 -1.1869 TD
[(plasmids)-230.9(encoding)-235.6(Cas9)-229.3(and)-234.7(target-specic)-232.7(gRNAs)]TJ
0 -1.1923 TD
[(to)-453.6(achieve)-452.3(editing)-449.3(in)-456(a)-450.6(mouse)-451.9(model)-450.2(of)]TJ
7.8454 0 0 6.9729 504.3967 194.1165 Tm
(70)Tj
10.4607 0 0 10.4607 517.7762 189.4676 Tm
[(and)-451.5(to)]TJ
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[(model)-287.6(cancer)-288.1(in)-288(the)-284(mouse)-289.3(liver.)]TJ
7.8454 0 0 6.9729 464.5983 181.644 Tm
(71)Tj
10.4607 0 0 10.4607 476.277 176.9952 Tm
[(Although)-287(these)]TJ
-16.1017 -1.1869 TD
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[(experiments)-366.8(provide)-366.1(a)-369.4(proof-of-concept)-368.4(validation)]TJ
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[(for)-421.7(gene)-424.7(editing)-422.2(of)-424.3(cells)-420.7(in)-423.5(an)-421.9(adult)-426.5(animal,)-421.2(this)]TJ
T*
[(approach)-280.2(is)-286.8(not)-278.6(directly)-283.1(applicable)-286.7(for)-280.8(clinical)-285.2(use.)]TJ
0 -1.1869 TD
[(Nevertheless,)-385.2(hydrodynamic)-388.8(delivery)-388.4(of)-386.4(CRISPR/)]TJ
0 -1.1923 TD
[(Cas9)-229.3(components)-221(is)-232.6(useful)-225.6(for)-226.7(the)-224.4(rapid)-227.7(generation)]TJ
T*
[(of)-180.5(mouse)-180.9(models)-181.9(for)-183.3(liver)-180.5(cancer)-185.1(and)-180.5(other)-182.5(diseases.)]TJ
/F15 1 Tf
1.1435 -2.39 TD
[(Lipid-mediated)-663.9(transfection.)]TJ
/F5 1 Tf
13.8038 0 TD
-.0119 Tc
[(On)-11.9(e)-610.5(o).3(f)-621.9(t)-.9(h)-11.9(e)-610.5(mo)-11.9(s)8.5(t)]TJ
-14.9473 -1.1869 TD
0 Tc
[(co)20.3(mm)19.5(on)-208.4(me)19.3(th)18.1(o)12.2(d)0(s)-211.1(f)0(o)20.1(r)-225(n)12.4(u)0(c)20.4(l)0(e)15.3(i)0(c)-211.1(a)0(c)18.1(i)13.9(d)-224.7(d)13.8(el)15.3(i)13.9(v)0(e)16.7(r)0(y)-209.7(t)11(o)-220.8(c)0(e)20.3(l)0(l)17(s)-219.5(in)]TJ
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[(a)-266.4(r)0(e)20.3(s)0(e)20.4(a)0(r)18.1(c)0(h)-256(c)0(o)20.2(n)0(t)18.1(e)0(x)16.7(t)-265.4(m)12.4(a)0(k)17.7(e)12.2(s)-268.2(u)0(s)20.6(e)-269.6(o)12.2(f)-263.2(li)17(pi)16.9(d)-268(n)12.4(an)17(o)12.2(p)0(a)18.4(r)0(t)19.1(i)0(c)16.5(l)13.9(es)]TJ
T*
[(o)12.2(r)-463.5(co)14.8(m)12.4(p)0(l)16.9(e)12.2(xe)16.7(s)13.6(.)-466.9(T)10.2(h)12.4(is)-454.8(ap)18.4(pr)16.4(o)12.2(a)0(c)18.1(h)-464.5(h)12.4(a)0(s)-453.3(e)0(n)19.2(t)11(er)14.8(e)12.2(d)-463.1(cl)16.5(i)13.9(n)0(i)15.5(c)0(a)18.1(l)]TJ
ET
41.839 753.902 508.082 .51025 re
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BT
/F14 1 Tf
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[(ADVAN)-9.5(CING)-337.1(CRIS)-9.4(PR)-336.1(THERA)-10.5(PEUTIC)-9.1(S)]TJ
/F13 1 Tf
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(445)Tj
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