ライフサイエンスコーパス: gRNA

1 gRNA expression through the commonly used U6 promoter re

2 targeting specificity of approximately 3,000 gRNAs across 30 therapeutically implicated loci.

3 lementation system involving bipartite HIV-1 gRNA, we observed that gRNA packaging is additionally de

4 To date, anti-HIV-1 gRNAs have been designed to account for off-target activ

5 is tests the ability of published anti-HIV-1 gRNAs to cleave publicly available patient-derived HIV-1

6 eletions in up to six genes by expressing 12 gRNAs from a single transcript.

7 ed duplicated screens using a library with 6 gRNAs per gene as providing the best trade-off.

8 Co-transfection of cells with Cas9, a gRNA specifying the genomic locus of interest, the donor

9 In addition, co-transformation of a gRNA plasmid and a donor DNA in cells constitutively exp

10 tion in the progression of editing through a gRNA; however, they have distinct roles and REMC is like

11 tent complexes, they can compete with active gRNAs for binding to Cas9.

12 fects on RBP16 gRNA and mRNA association and gRNA-containing ribonucleoprotein complex (gRNP) formati

13 le-stranded determinants for association and gRNA-directed cleavage.

14 An appropriate combination of Cas9 and gRNA allows targeting of essential and nonessential gene

15 though specific interactions between Gag and gRNA have been demonstrated previously, where and when t

16 They also bind gRNAs and gRNA/pre-mRNA hybrid RNAs with similar affinities and as

17 se pairing between their respective mRNA and gRNA cargo and regulatory REH2 and (H2)F1 subunits of th

18 ther virion proportions of Gag, Gag-Pol, and gRNA were determined by sampling (that is, if they refle

19 To this end, MoMLV Gag, Gag-Pol, and gRNA were expressed separately or together in various ra

20 and NC domains affects virus replication and gRNA packaging efficiency.

21 e the production of longer viral sgmRNAs and gRNA.

22 e distance between the modification site and gRNA target site was a significant parameter affecting t

23 SPR binding specificity at gRNA-specific and gRNA-independent sites for two guide RNAs.

24 e show high-level concentration of virus and gRNA in lymph nodes after intramuscular inoculation of m

25 s and multiplexed production of proteins and gRNAs from a single transcript in human cells.

26 twice as effective in inducing mutations as gRNAs expressed from individual RNA polymerase III promo

27 ne genome-wide CRISPR binding specificity at gRNA-specific and gRNA-independent sites for two guide R

28 n designing screens and constructing bespoke gRNA libraries.

29 ysis suggests that mismatch position between gRNA seed and target DNA is an important determinant of

30 They also bind gRNAs and gRNA/pre-mRNA hybrid RNAs with similar affinit

31 The helix bundle binds gRNA, causing denser packing of RNA in its proximity, wh

32 stable 200 kDa particle that directly binds gRNAs.

33 alpha-helix II, previously implicated in BLV gRNA packaging, reduces NA binding affinity.

34 e web interface of pgRNAFinder contains both gRNA search and scoring system.

35 Although both gRNAs and mRNAs are associated with the RESC, their meta

36 viruses) supported translationin vitro, but gRNA did not accumulate to detectable levels in protopla

37 -base Guanine constraint commonly imposed by gRNA expression systems has little effect on overall cle

38 y second-generation genome-scale CRISPR-Cas9 gRNA library and applied it to fitness screens in five h

39 r-Like Effectors Nucleases (TALENs) and Cas9-gRNA allows genetic modifications to be made more effici

40 Cs mediated by 15 pairs of reTALENs and Cas9-gRNA targeting CCR5 and optimized ssODN design in conjun

41 ort biolistic delivery of pre-assembled Cas9-gRNA ribonucleoproteins into maize embryo cells and rege

42 We found Cas9-gRNA achieved 7-8x higher non-homologous end joining eff

43 rom either failure to form a functional Cas9-gRNA complex or inability to recognize targets in vivo.

44 previously uncharacterized features of Cas9-gRNA complex formation.

45 ro understanding of the complexities of Cas9-gRNA interaction and cleavage beyond the general paradig

46 nome modifications were specific to the Cas9-gRNA cleavage sites and consisted of small deletions or

47 The Cas9-gRNA system was also successfully applied to make a dire

48 delivering a CRISPR plasmid expressing Cas9/gRNA and a single-stranded oligodeoxynucleotide HDR dono

49 ared mutational tolerance for a set of Cas9::gRNA complexes in vitro and in vivo (in Saccharomyces ce

50 HIV-1 reading frame, while destroying CRISPR gRNA homology.

51 avage site were major contributors to CRISPR gRNA resistance.

52 and exhibited both precleaved and full-cycle gRNA-mediated U-insertion and U-deletion in vitro activi

53 Systemic delivery of dCas9/gRNA by adeno-associated virus led to reductions in path

54 Here, we define rules that determine gRNA effectiveness for transcriptional repression in Sac

55 the two conserved GRPE stem loops diminished gRNA packaging and infectivity >50-fold, while deleting

56 he RESC, their metabolic fates are distinct: gRNAs are degraded in an editing-dependent process, wher

57 d provide guidelines for designing effective gRNAs, which consider chromatin state and position relat

58 tion, the off-target effect of an engineered gRNA-Cas9 was found on an imperfectly paired genomic sit

59 The engineered gRNAs were shown to direct the Cas9 nuclease for precise

60 Not every gRNA elicits cleavage and the mechanisms that govern gRN

61 ticle production was not disrupted by excess gRNA expression.

62 endogenous genes using H1 promoter-expressed gRNAs, which can be used to target both AN19NGG and GN19

63 ized that Gag nuclear entry might facilitate gRNA packaging.

64 rinciple use of multiplexed ribozyme flanked gRNAs to induce mutations in vivo in Drosophila melanoga

65 ification of a protein complex essential for gRNA stability.

66 e in size, and encode between three and four gRNAs.

67 ategies that enable expression of functional gRNAs from RNA polymerase II promoters and multiplexed p

68 ag-Pol proportions differ from those for Gag/gRNA.

69 cits cleavage and the mechanisms that govern gRNA activity have not been resolved.

70 Comparing near-identical gRNA sequences with different in vitro activities reveal

71 imple and quick screening method to identify gRNA candidates for targeting HIV provirus in astrocytes

72 ting machinery, with additional functions in gRNA and mRNA stabilization.

73 cell lines indicate that TbRND functions in gRNA metabolism in vivo.

74 These data indicate that REH1 is involved in gRNA displacement either directly by unwinding the gRNA/

75 the nucleocapsid (NC) domain is involved in gRNA packaging and displays robust nucleic acid (NA) bin

76 First, TbRND depletion results in gRNA tails extended by 2-3 nucleotides on average.

77 different retroviral Gag proteins influence gRNA packaging, highlighting variations and similarities

78 le patient-derived HIV-1 sequences to inform gRNA design and provides basic computational tools to re

79 eveal that both genomic context and internal gRNA interactions can interfere with Cas9-mediated cleav

80 nt in vitro activities reveals that internal gRNA interactions reduce cleavage.

81 irectly convert guide RNA (gRNA) inputs into gRNA outputs, enabling the gates to be 'wired' together.

82 ere efficiently and precisely processed into gRNAs with desired 5' targeting sequences in vivo, which

83 The kctd10 gRNA, designed against an essential functional region of

84 isolated this mRNP from mitochondria lacking gRNA-bound RNP (gRNP) subcomplexes and identified REH2-a

85 rget) is not clearly superior to full-length gRNAs (20 nt of complementarity), as truncated gRNAs are

86 er, the mechanism(s) that selects and limits gRNAs for packaging remains uncertain.

87 By microinjecting gRNA, hCas9 mRNA and single-stranded donor oligonucleoti

88 RNA unwinding activity in vitro with a model gRNA-mRNA duplex.

89 To avoid DCK gene modification, gRNA resistant DCK cDNA was created by the introduction

90 ive virus particles containing 20 times more gRNA.

91 s achieved with microinjection of Cas9 mRNA, gRNA and single strand oligonucleotide DNA (ssDNA) into

92 serve as scaffolds for the assembly of mRNA-gRNA hybrids and RECC.

93 se the common use of artificially tight mRNA:gRNA base pairing precludes alternate alignments.

94 and all-in-one delivery of Cas9 and multiple gRNA expression cassettes with AAV vectors.

95 ribe a simple method for expressing multiple gRNAs bearing any 5' end nucleotide, which gives dimeric

96 By efficiently co-expressing multiple gRNAs that target different genomic sites, the polycistr

97 s strategy allows the expression of multiple gRNAs for synergistic transcription activation of follis

98 , upon simultaneous introduction of multiple gRNAs, can effect multiplex editing of target loci.

99 Here we show that single or multiple gRNAs can direct dCas9 fused to a VP64 transcriptional a

100 editing "block." Many mRNAs require multiple gRNAs; the observed overall 3' to 5' polarity of editing

101 Here, we report that multiple gRNAs linked with self-cleaving ribozymes and/or tRNA co

102 plexed gRNAs and that, with four multiplexed gRNAs, a mosquito species could potentially be suppresse

103 exponentially with the number of multiplexed gRNAs and that, with four multiplexed gRNAs, a mosquito

104 primary uridylation of approximately 800 nt gRNA precursors, their processive degradation to a matur

105 loped a general strategy to produce numerous gRNAs from a single polycistronic gene.

106 virus-like particles even in the absence of gRNA binding, whether viral RNA trafficking plays an act

107 llows for the rapid and efficient cloning of gRNA pairs into expression vectors.

108 The determinants of gRNA proportions were addressed by manipulating the amou

109 e RNA (gRNA) scaffold and the development of gRNA on-target prediction algorithms, have since been ma

110 the improvements, and examined the effect of gRNA scaffold, number of gRNAs per gene and number of re

111 king and also incorporates reduced levels of gRNA into virus particles compared to those in wild-type

112 An upper limit of gRNA incorporation was observed, and particle production

113 We observe a fixed pattern of gRNA organization among all viral particles, with the ma

114 on, our results indicate that persistence of gRNA does not result from continuing viral replication.

115 vectors as well as long-term persistence of gRNA in the lymph nodes.

116 To determine if the persistence of gRNA was due to ongoing viral replication, we developed

117 s required for the nucleolytic processing of gRNA, rRNA, and mRNA precursors.

118 ractions stabilize the tertiary structure of gRNA within the virion, which could further provide a ro

119 at a step prior to primary transcription of gRNA into mRNA.

120 To extend the understanding of gRNA::target homology requirements, we compared mutation

121 bunit of the 3' processome in uridylation of gRNA precursors and mature guide RNAs.

122 is limited while tissue-specific delivery of gRNAs and Cas9 is desired.

123 structure of GuideScan enables the design of gRNAs that are more specific than those designed by exis

124 st, and use it to analyze fitness effects of gRNAs under 18 small molecule treatments.

125 , GRBC1 and GRBC2, led to the elimination of gRNAs, thus inhibiting mRNA editing.

126 (RRE), which regulates the nuclear export of gRNAs and other intron-retaining viral RNAs.

127 In Trypanosoma brucei, the vast majority of gRNAs are transcribed from minicircles, which are approx

128 tion rate but rather increased the number of gRNAs available for translation.

129 mined the effect of gRNA scaffold, number of gRNAs per gene and number of replicates on screen perfor

130 particle responsible for the recognition of gRNAs and pre-mRNA substrates, editing intermediates, an

131 To identify the complete set of gRNAs necessary for mRNA editing in T. brucei, we used I

132 We report a near complete set of gRNAs needed to direct the editing of the mRNAs.

133 's presence does not affect the stability of gRNAs and rRNAs, while transcript-specific uridylylation

134 A variety of gRNAs were tested with variant libraries based on four d

135 PgRNAFinder offers gRNA design functionality for 8 vertebrate genomes.

136 most of the existing computational tools on gRNA design are restricted to small deletions.

137 One gRNA identified in this screen outperformed the most pro

138 tissue-specific expression of more than one gRNAs for multiplex gene editing from a single pol II pr

139 c changes in association kinetics when other gRNA-target mismatches are present.

140 editing mediated by two or more overlapping gRNAs but has no effect on editing within a single block

141 ental approaches to generate pools of paired gRNA vectors prevents these applications from being scal

142 of guide RNAs (gRNAs) for single- and paired-gRNA genome-wide screens.

143 system and two specificity-enhancing paired-gRNA systems: Cas9 D10A nickases (Cas9n) and dimeric RNA

144 ers to design single or distance-free paired-gRNA sequences.

145 Unlike Gag-Pol, gRNA incorporation was saturable.

146 entially modulated by this strong protective gRNA that rescued yeast from alphaSyn toxicity when over

147 omain of either Gag or Gag-Pol could provide gRNA packaging functions equally well.

148 we used Illumina deep sequencing of purified gRNAs from the procyclic stage.

149 rg methylation has distinct effects on RBP16 gRNA and mRNA association and gRNA-containing ribonucleo

150 Also, the NLS sequence restored gRNA packaging to nearly wild-type levels in viruses con

151 virus mRNA and negative-strand genomic RNA (gRNA) accumulated to high levels at 8 h after infection

152 he maturation protein binds the genomic RNA (gRNA) and is required for attachment of the phage to the

153 ves a recognition event between genomic RNA (gRNA) and one or more domains in Gag.

154 l expression of the CP from the genomic RNA (gRNA) both in vitro and in vivo An absence of extensive

155 -Pol polyproteins plus a single genomic RNA (gRNA) dimer.

156 rs showed persistence of vector genomic RNA (gRNA) for at least 60 days in lymph nodes in the absence

157 s to selective packaging of the genomic RNA (gRNA) into virions.

158 for annealing tRNA(Lys3) to the genomic RNA (gRNA) primer binding site (PBS).

159 The packaging of retroviral genomic RNA (gRNA) requires cis-acting elements within the RNA and tr

160 Selective packaging of HIV-1 genomic RNA (gRNA) requires the presence of a cis-acting RNA element

161 around its 4,217 nucleotides of genomic RNA (gRNA).

162 of longer sgmRNAs and the viral genomic RNA (gRNA).

163 eocapsid protein (N) with viral genomic RNA (gRNA).

164 infectivity of transfected MHV genomic RNA (gRNA).

165 most likely due to alternate mRNA:guide RNA (gRNA) alignment forming a hyphenated anchor; its having

166 ckout lines by simply injecting a guide RNA (gRNA) and Cas9 mRNA into one-cell stage embryos.

167 combinatorial mismatches between guide RNA (gRNA) and target nucleotides, both in the seed and in mo

168 genome of cell lines to evaluate guide RNA (gRNA) efficiency, safety, and toxicity.

169 ansgenic Cas9 lines and versatile guide RNA (gRNA) expression plasmids.

170 PR system to function with custom guide RNA (gRNA) in human cells.

171 f NOR gates that directly convert guide RNA (gRNA) inputs into gRNA outputs, enabling the gates to be

172 Using a genome-wide guide RNA (gRNA) library, we found that targeting Nek7 rescued macr

173 fication in the expression of the guide RNA (gRNA) required for targeting that greatly expands the ta

174 nts, such as modifications to the guide RNA (gRNA) scaffold and the development of gRNA on-target pre

175 high efficiency with a variety of guide RNA (gRNA) spacer lengths.

176 We include guidelines for guide RNA (gRNA) target design, embryo injection and hatching, germ

177 s, we designed a disease-specific guide RNA (gRNA) targeting the R124H mutation of TGFBI, which cause

178 This system uses a small guide RNA (gRNA) to direct Cas9 endonuclease to a specific DNA site

179 clease (Cas9) system depends on a guide RNA (gRNA) to specify its target.

180 diting method is comprised of the guide RNA (gRNA) to target a specific DNA sequence for cleavage and

181 sses of pre-mRNA polyadenylation, guide RNA (gRNA) uridylylation and annealing to mRNA, and editing r

182 UTases) are known: RET1 catalyzes guide RNA (gRNA) uridylylation, RET2 executes U insertion mRNA edit

183 Streptococcus pyogenes Cas9-guide RNA (gRNA) was successfully applied to generate targeted muta

184 ing the length of Cas9-associated guide RNA (gRNA) we were able to control Cas9 nuclease activity and

185 doxycycline and transfection with guide RNA (gRNA), donor DNA and piggyBac transposase resulted in ef

186 designer genome targeting CRISPR guide RNA (gRNA), show robust and specific RNA-guided endonuclease

187 cycles of three catalytic steps: guide RNA (gRNA)-directed cleavage, insertion or deletion of uridyl

188 nding on their position along the guide RNA (gRNA)-DNA interface.

189 a library of 23,409 barcoded dual guide-RNA (gRNA) combinations and then perform a high-throughput po

190 By delivering Cas9 and guide-RNA (gRNA) with retro- or lenti-virus to IgM(+) mouse B cells

191 Pol capsid proteins as well as genomic RNAs (gRNAs) packaged by Gag into virions undergoing assembly

192 ing system architecture in which guide RNAs (gRNAs) are multiplexed, increasing the effective homing

193 Over 1200 small guide RNAs (gRNAs) are predicted to be responsible for directing the

194 Short guide RNAs (gRNAs) can direct catalytically inactive CRISPR-associat

195 are directed to genomic loci by guide RNAs (gRNAs) containing 20 nucleotides that are complementary

196 tion of this methodology is that guide RNAs (gRNAs) for CRISPR-TFs can only be expressed from RNA pol

197 an produces high-density sets of guide RNAs (gRNAs) for single- and paired-gRNA genome-wide screens.

198 wever, the features of effective guide RNAs (gRNAs) in different organisms have not been well charact

199 hough complexes between Cas9 and guide RNAs (gRNAs) offer remarkable specificity and versatility for

200 web service to help users design guide RNAs (gRNAs) optimized for specificity.

201 ate pre-edited mRNAs and cognate guide RNAs (gRNAs) represents the first step in the reaction cycle,

202 We designed 87,897 guide RNAs (gRNAs) targeting 19,150 mouse protein-coding genes and u

203 computationally designed unique guide RNAs (gRNAs) targeting all VACV genes will be valuable for the

204 nd directed by hundreds of small guide RNAs (gRNAs) that base pair with mRNA.

205 ltaneously using two CRISPR/Cas9 guide RNAs (gRNAs) that depend on PAM sites generated by SNP alleles

206 However, guide RNAs (gRNAs) that direct U-insertion/deletion mRNA editing in

207 on's disease (PD), we identified guide RNAs (gRNAs) that modulate transcriptional networks and protec

208 cluding the 3' oligo(U) tails of guide RNAs (gRNAs) that provide the sequence information for RNA edi

209 The CRISPR/Cas system uses guide RNAs (gRNAs) to direct sequence-specific DNA cleavage.

210 s strictly on the binding of two guide RNAs (gRNAs) to DNA with a defined spacing and orientation sub

211 -based system that uses pairs of guide RNAs (gRNAs) to program thousands of kilobase-scale deletions

212 ey components of CRISPR/Cas9 are guide RNAs (gRNAs) which determine specific sequence targeting of DN

213 Three guide RNAs (gRNAs) with a 20-22-nt seed region were designed to pair

214 ein 9 (Cas9), including specific guide RNAs (gRNAs), can excise integrated human immunodeficiency vir

215 ion for editing resides in small guide RNAs (gRNAs), which form anchor duplexes just downstream of an

216 re co-expression of two distinct guide RNAs (gRNAs).

217 ny genomic locus using so called guide RNAs (gRNAs).

218 al, and ligation are directed by guide RNAs (gRNAs).

219 simultaneously express multiple guide RNAs (gRNAs).

220 with the co-delivery of multiple guide RNAs (gRNAs).

221 nce to six different CRISPR/Cas9 guide RNAs (gRNAs).

222 emonstrate differential activity of the same gRNA expressed from different U6 snRNA promoters, with t

223 ter users input query sequences, it searches gRNA by 3' protospacer-adjacent motif (PAM), and possibl

224 As expected, sequencing gRNA pairs before and after selection confirmed that all

225 constructed logic circuits with up to seven gRNAs, including repression cascades with up to seven la

226 oceed through numerous paths within a single gRNA and that non-linear modifications are essential, ge

227 RFNs guided by a single gRNA generally induce lower levels of unwanted mutations

228 Both single gRNA/WT hCas9 and double nicking set-ups were effective.

229 how that targeting of these loci with single gRNAs leads to efficient and widespread methylation of t

230 CT-Finder accommodates the original single-gRNA Cas9 system and two specificity-enhancing paired-gR

231 t, the donor plasmid and a cassette-specific gRNA triggers the insertion of the tag by a homology-ind

232 ession of both a multiplex of HIV-1-specific gRNAs and Cas9 in cells results in the modification and/

233 RNA-Cas9 targeting specificity, and specific gRNAs could be designed to target more than 90% of rice

234 We also find that the best region to target gRNAs is between the transcription start site (TSS) and

235 IRES region in vitro by use of both the TCV gRNA and reporter constructs did not reveal any sequence

236 lving bipartite HIV-1 gRNA, we observed that gRNA packaging is additionally dependent on a cis-acting

237 We propose that gRNA is selectively packaged because binding to Psi nucl

238 Finally, we show that gRNA binding proteins co-purify with TbRND.

239 Furthermore, we found that gRNAs represent only a subset of small mitochondrial RNA

240 The gRNA-binding complex (GRBC) interacts with gRNA processi

241 cture, at 7-A resolution, reveals A2 and the gRNA.

242 ing capability is largely constrained by the gRNA-expressing device.

243 in the double-stranded DNA identified by the gRNA.

244 y suggesting that the IRES was active in the gRNA invivo Since the TCV CP also serves as the viral si

245 Chemical modification of the gRNA and donor DNA has great potential for improving the

246 The tolerance of the gRNA and donor DNA to chemical modifications has the pot

247 Both strands of the gRNA gene are transcribed into sense and antisense precu

248 city increase in vivo with truncation of the gRNA homology regions.

249 hich Psi promotes selective packaging of the gRNA is not well understood.

250 uctures in the 3' untranslated region of the gRNA, contains the promoter for negative-strand synthesi

251 arget DNA is an important determinant of the gRNA-Cas9 targeting specificity, and specific gRNAs coul

252 ation produces an increased abundance of the gRNA/edited mRNA duplex for the first editing block of t

253 The observed effects on the gRNA population are specific as rRNAs, which are also 3'

254 rly stage of capsid formation to promote the gRNA condensation required for genome packaging.

255 In addition, we demonstrate that the gRNA and donor DNA can be directly conjugated together i

256 This coat protein dimer binds to the gRNA and interacts with the buried alpha-region of A2, s

257 on, the maturation protein, A2, binds to the gRNA and is required for adsorption to the F-pilus.

258 isplacement either directly by unwinding the gRNA/edited mRNA duplex or indirectly, to allow the 5' a

259 The CRISPR screening was repeated using the gRNA resistant DCK, and loss of SLC29A was identified as

260 eintegration of the region demarcated by the gRNAs in the vicinity of the edited locus.

261 Finally, the gRNAs linked by the self-cleaving ribozymes and tRNA cou

262 In this report, we demonstrate that the gRNAs of Cas9 and Cpf1, and donor DNA can be chemically

263 nd used a lentiviral vector to express these gRNAs in ESCs that constitutively express Cas9.

264 Moreover, these gRNAs and Cas9 protein were successfully tested on HIV l

265 gether into one molecule, and show that this gRNA-donor DNA conjugate is three times better at transf

266 Even though gRNAs containing these structures do not yield cleavage-

267 l nucleocapsid (NC) and the ratios of Gag to gRNA.

268 We show that RET1 adds U tails to gRNAs, rRNAs, and select mRNAs and contributes U's into

269 sults in a substantial decrease in the total gRNA population and a consequent inhibition of RNA editi

270 constitutive Cas9 expression and a transient gRNA cassette, we show that targeted double-strand break

271 t synthetic genes with tandemly arrayed tRNA-gRNA architecture were efficiently and precisely process

272 ferent genomic sites, the polycistronic tRNA-gRNA gene (PTG) strategy enables multiplex gene editing

273 NAs (20 nt of complementarity), as truncated gRNAs are generally less potent against both mismatched

274 in human cells, the specificity of truncated gRNAs (18 nt of complementarity to the target) is not cl

275 In addition, use of truncated gRNAs can further reduce off-target effects induced by p

276 Our study first demonstrates that truncated gRNAs to 18 complementary nucleotides and Cas9 nucleases

277 Here we report that truncated gRNAs, with shorter regions of target complementarity <2

278 contains the tag of interest flanked by two gRNA recognition sites that allow excision of the tag fr

279 n, possibly explaining the limitation of two gRNAs per virion.

280 eus Cas9, and we further package it with two gRNAs in a single functional adeno-associated virus (AAV

281 R, with cationic polymers, than unconjugated gRNA and donor DNA.

282 pute a genome-wide resource of ~190 K unique gRNAs targeting ~40.5% of human exons.

283 ndirectly, to allow the 5' adjacent upstream gRNA to form an anchor duplex with the edited mRNA to in

284 multiple target loci across treatments using gRNA libraries allows us to determine generalizable feat

285 Various gRNAs were screened for their efficiencies against HIV p

286 oteins that is required to package the viral gRNA in its dominant conformation.

287 , early translation of the CP from the viral gRNA is likely important for countering host defenses.

288 aging signal psi, at the 5' end of the viral gRNA, binds to Gag through interactions with basic resid

289 virus genome RNA (cRNA) and influenza virus gRNA were drastically suppressed.

290 are consistent with initial trapping of VSV gRNA largely in lymph node macrophages and subsequent pe

291 Our results show that VSV gRNA persists long-term in the lymph nodes while VSV mRN

292 target editing activity had been varied when gRNAs was truncated, higher at Site Two (tF7-2 vs. F7-2,

293 support a model for Cas9 specificity wherein gRNA-DNA mismatches at PAM-distal bases modulate differe

294 ether, our findings suggest a model in which gRNA is derived from the 5' extremity of a primary molec

295 rmine generalizable features associated with gRNA efficacy.

296 em possess 3' U-tails, which correlates with gRNA's enrichment in the RESC.

297 12 nt and an approximately 15-bp duplex with gRNA to direct the cleavage site.

298 e gRNA-binding complex (GRBC) interacts with gRNA processing, editing, and polyadenylation machinerie

299 ructural basis of the interaction of MA with gRNA, host transport factors and membrane phospholipids.

300 afficking of the retroviral Gag protein with gRNA incorporation.