ライフサイエンスコーパス: guide RNA

1 that precede the pairing of target DNA with guide RNA.

2 ectly measure the stability of both Cas9 and guide RNA.

3 from enhancer regions and with an individual guide RNA.

4 activity to the previously published single guide RNA.

5 iviral vectors that express Cre and a single-guide RNA.

6 acent motif (PAM) and complementarity to the guide RNA.

7 s to the gene targeted by the co-transfected guide RNA.

8 tivity but that this activity depends on the guide RNA.

9 irs allowing initial target interrogation by guide RNA.

10 od rather than plasmids that encode Cas9 and guide RNAs.

11 patients with CRISPR/Cas9 using one pair of guide RNAs.

12 in uridylation of gRNA precursors and mature guide RNAs.

13 -specific and gRNA-independent sites for two guide RNAs.

14 can easily be scaled up to process multiple guide RNAs.

15 modification characteristic of ribosomal and guide RNAs.

16 complexes to target loci by modified single guide RNAs.

17 on or deletion, and ligation as specified by guide RNAs.

18 n vivo); 16 websites used to generate single-guide RNA; 4 websites for off-target effects; and 382 ar

19 estabilization is mediated, in part, by Cas9/guide RNA affinity for unpaired segments of nontarget st

20 undergoes major conformational changes upon guide RNA and DNA binding.

21 get through base pairing between a synthetic guide RNA and DNA, outperforms zinc-finger proteins and

22 le-stranded DNA targets complementary to the guide RNA and has been applied to programmable genome ed

23 eak at DNA target sites complementary to the guide RNA and has been harnessed for the development of

24 he largest Cas9 orthologs, in complex with a guide RNA and its PAM-containing DNA targets.

25 coccus sp. Cpf1 (AsCpf1) in complex with the guide RNA and its target DNA at 2.8 A resolution.

26 o method for the simultaneous examination of guide RNA and protospacer adjacent motif (PAM) requireme

27 The combination of Cas9, guide RNA and repair template DNA can induce precise gen

28 Both nucleases, in the presence of a guide RNA and repairing DNA template flanked by homology

29 en by complementary base pairing between the guide RNA and target DNA, Cas9-DNA interactions, and ass

30 anism driven by the interactions between the guide RNA and the 14th-17th nucleotide region of the tar

31 e expressing Cas9 and the other expressing a guide RNA and the donor DNA, into newborn mice with a pa

32 of at least one mismatch between the single guide RNA and the non-target gene sequences.

33 dified by U-insertion/deletion editing while guide RNAs and rRNAs are U-tailed.

34 repeats (CRISPR)-associated protein 9 (Cas9) guide RNAs and target interrogation are not well defined

35 kage between ADAR's catalytic domain and the guide RNA, and by introducing a mutation in the catalyti

36 rary that consisted of around 123,000 single-guide RNAs, and profiled genes whose loss in tumour cell

37 Finally, we applied a dual short guide RNA approach to effectively delete an important co

38 ing a fluorescent protein, Cas9, and a small guide RNA are used to mimic nonsense PTEN mutations from

39 Biolistic delivery of guide RNAs (as RNA molecules) directly into immature emb

40 cally stable 5-(E)-vinylphosphonate (5-E-VP) guide RNA at 2.5-A resolution.

41 least 4.8 kb long can be inserted in CRISPR guide RNA at multiple points, allowing the construction

42 f Cas9 endonuclease for DNA recognition upon guide RNA binding occurs by an unknown mechanism.

43 The results show that Cas9/guide RNA binding to PAM favors separation of a few PAM-

44 structure of AcrIIA4 in complex with single-guide RNA-bound SpyCas9, thereby establishing that AcrII

45 leases target specific DNA sequences using a guide RNA but also require recognition of a protospacer

46 ed a method to photocage the activity of the guide RNA called "CRISPR-plus" (CRISPR-precise light-med

47 Together with single-guide RNAs, Cas9 also functions as a powerful genome eng

48 The iPSCs were transduced with CRISPR guide RNAs, Cas9 endonuclease, and a donor homology temp

49 -specific Cas9 cassette together with single-guide RNA cassettes and, in one approach, a dystrophin h

50 Very few microRNAs are known to guide RNA cleavage.

51 NA targeting, in vitro data have shown crRNA-guided RNA cleavage.

52 ining a randomized PAM as a function of Cas9-guide RNA complex concentration.

53 uide RNA stability and that the nuclear Cas9-guide RNA complex levels limit the targeting efficiency.

54 onic lipid-mediated in vivo delivery of Cas9-guide RNA complexes can ameliorate hearing loss in a mou

55 Cas9-guide RNA complexes recognize 20-base-pair sequences in

56 Cas9-guide RNA complexes recognize and cleave double-stranded

57 Binding specificity of Cas9-guide RNA complexes to DNA is important for genome-engin

58 tric and biochemical assays, we studied Cas9/guide RNA complexes with model DNA substrates that mimic

59 associated 9 from Staphylococcus aureus) and guide RNA constructs into an adeno-associated virus vect

60 The CjCas9 guide RNA contains a triple-helix structure, which is di

61 We also observed that multiple single-guide RNAs could be expressed in the same transcript, re

62 We also showed that multiple guide RNAs could target the dCas9-DNMT3A construct to mu

63 ive Cas9 protein together with gene-specific guide RNAs (CRISPR interference).

64 five different Cas proteins and a 61-nt-long guide RNA (crRNA).

65 Furthermore, using a single guide RNA, dCas9-SunTag-DNMT3A is able to methylate a 4.

66 of Cas9 together with JCPyV-specific single-guide RNA delivered prior to or after JCPyV infection.

67 NA sequencing reveals that recCas9 catalyzes guide RNA-dependent recombination in human cells with an

68 potential off-targets that enables improved guide RNA design and more accurate prediction of Cas9 bi

69 mismatch with implications for mechanism and guide-RNA design.

70 king site with a DNA polymerase to label the guide RNA-determined target sequences.

71 nces similar to those targeted by individual guide RNAs did not reveal significant off-target effects

72 he two nuclease domains of Cas9, to create a guide RNA-directed DNA nick in the context of an in vitr

73 M-proximal region during early steps of Cas9/guide RNA-DNA complex formation, thus additionally desta

74 termediates on the pathway to the final Cas9/guide RNA-DNA complex.

75 t retain the ability to be programmed with a guide RNA, do not induce dsDNA breaks, and mediate the d

76 Multiplexing of several guide RNAs does not increase the efficiency of methylati

77 livery of recombinant Cas9 protein and short guide RNA, driving efficient gene targeting in a non-int

78 gle-subunit C2c2 protein functions as an RNA-guided RNA endonuclease (RNase).

79 Finally, GUIDE-seq revealed that truncated guide RNAs exhibit substantially reduced RGN-induced, of

80 e a cardiac phenotype, irrespective of short guide RNA exposure or the level of Cas9 expression.

81 We packaged SaCas9 and its single guide RNA expression cassette into a single AAV vector a

82 uding tissue-specific promoters, multiplexed guide RNA expression, and effector domain fusions to SpC

83 Collectively, our results establish guide RNA features that drive DNA targeting by Cas9 and

84 ISPR)-Cas9 endonucleases coupled with paired guide RNAs flanking the mutated Dmd exon23 resulted in e

85 re able to rapidly identify highly effective guide RNAs; focusing germ line-based experiments only on

86 SPR/Cas9 endonuclease system, comprised of a guide RNA for the recognition of a DNA target and the Ca

87 stingly, all drug resistant clones contained guide RNAs for DCK.

88 cores for all guides, thereby offering rapid guide RNA generation and selection.

89 tion of knockout lines by simply injecting a guide RNA (gRNA) and Cas9 mRNA into one-cell stage embry

90 effects of combinatorial mismatches between guide RNA (gRNA) and target nucleotides, both in the see

91 nome or the genome of cell lines to evaluate guide RNA (gRNA) efficiency, safety, and toxicity.

92 l library of NOR gates that directly convert guide RNA (gRNA) inputs into gRNA outputs, enabling the

93 Using a genome-wide guide RNA (gRNA) library, we found that targeting Nek7 r

94 s improvements, such as modifications to the guide RNA (gRNA) scaffold and the development of gRNA on

95 get DNA at high efficiency with a variety of guide RNA (gRNA) spacer lengths.

96 We include guidelines for guide RNA (gRNA) target design, embryo injection and hat

97 atient cells, we designed a disease-specific guide RNA (gRNA) targeting the R124H mutation of TGFBI,

98 This system uses a small guide RNA (gRNA) to direct Cas9 endonuclease to a specif

99 rotein 9 nuclease (Cas9) system depends on a guide RNA (gRNA) to specify its target.

100 Cas9 gene editing method is comprised of the guide RNA (gRNA) to target a specific DNA sequence for c

101 Streptococcus pyogenes Cas9-guide RNA (gRNA) was successfully applied to generate ta

102 at by altering the length of Cas9-associated guide RNA (gRNA) we were able to control Cas9 nuclease a

103 tment with doxycycline and transfection with guide RNA (gRNA), donor DNA and piggyBac transposase res

104 to create a library of 23,409 barcoded dual guide-RNA (gRNA) combinations and then perform a high-th

105 By delivering Cas9 and guide-RNA (gRNA) with retro- or lenti-virus to IgM(+) mo

106 xplore a homing system architecture in which guide RNAs (gRNAs) are multiplexed, increasing the effec

107 GuideScan produces high-density sets of guide RNAs (gRNAs) for single- and paired-gRNA genome-wi

108 ll lines; however, the features of effective guide RNAs (gRNAs) in different organisms have not been

109 Although complexes between Cas9 and guide RNAs (gRNAs) offer remarkable specificity and vers

110 Finder is a web service to help users design guide RNAs (gRNAs) optimized for specificity.

111 set of 8,964 computationally designed unique guide RNAs (gRNAs) targeting all VACV genes will be valu

112 lex (RECC) and directed by hundreds of small guide RNAs (gRNAs) that base pair with mRNA.

113 nciple, simultaneously using two CRISPR/Cas9 guide RNAs (gRNAs) that depend on PAM sites generated by

114 However, guide RNAs (gRNAs) that direct U-insertion/deletion mRNA

115 l of Parkinson's disease (PD), we identified guide RNAs (gRNAs) that modulate transcriptional network

116 The CRISPR/Cas system uses guide RNAs (gRNAs) to direct sequence-specific DNA cleav

117 ibe a CRISPR-based system that uses pairs of guide RNAs (gRNAs) to program thousands of kilobase-scal

118 The key components of CRISPR/Cas9 are guide RNAs (gRNAs) which determine specific sequence tar

119 ociated protein 9 (Cas9), including specific guide RNAs (gRNAs), can excise integrated human immunode

120 oped resistance to six different CRISPR/Cas9 guide RNAs (gRNAs).

121 tions--require co-expression of two distinct guide RNAs (gRNAs).

122 to almost any genomic locus using so called guide RNAs (gRNAs).

123 g enzymes to simultaneously express multiple guide RNAs (gRNAs).

124 es feasible with the co-delivery of multiple guide RNAs (gRNAs).

125 A homing guide RNA (hgRNA) scaffold directs the Cas9-hgRNA comple

126 is shown to functionally replace the natural guide RNA in the CRISPR-Cas9 nuclease system and to medi

127 xpression of Cas9 and the specific targeting guide RNAs in HIV-1-eradicated T-cells protected them ag

128 geted all pairs of 73 cancer genes with dual guide RNAs in three cell lines, comprising 141,912 tests

129 They guide RNA-induced silencing complexes to complementary t

130 MicroRNAs (miRNAs) guide RNA-induced silencing complexes to target RNAs bas

131 mRNA duplexes for regulation of translation, guide RNA interactions with target RNA for post-transcri

132 MicroRNA (miRNA) biogenesis and miRNA-guided RNA interference (RNAi) are essential for gene ex

133 njection of Cas9 mRNA, eGFP mRNA, and single guide RNAs into fertilized eggs.

134 on of recombinant Cas9 protein and synthetic guide RNAs into mouse zygotes has been shown to facilita

135 d specificity by considering the invasion of guide RNAs into targeted DNA duplex.

136 This segment of the guide RNA is essential for Cas9 to form a DNA recognitio

137 iction model strongly depends on whether the guide RNA is expressed from a U6 promoter or transcribed

138 whether the target gene is essential or the guide RNA is ineffective.

139 d virus serotype 9 to deliver a single short guide RNA is target dependent.

140 ports numerous modifications that can enable guide RNA labeling for use in imaging and mechanistic in

141 Custom- or ready-made guide RNA libraries are constructed and packaged into le

142 nuclease has been combined with genome-scale guide RNA libraries for unbiased, phenotypic screening.

143 named "CRISPR EATING" for generating complex guide RNA libraries suitable for CRISPR/Cas9-based appli

144 GuideScan software for the design of CRISPR guide RNA libraries that can be used to edit coding and

145 interspaced palindromic repeat (CRISPR)-Cas9 guide RNA libraries to perform in situ saturating mutage

146 ) activator constructs and lentiviral single guide RNA libraries to target DNase I hypersensitive sit

147 Using pooled lentiviral single-guide RNA libraries, we conducted a genome-wide loss-of-

148 cale screening by combining Cas9 with pooled guide RNA libraries.

149 system, we constructed a genome-wide single-guide RNA library to screen for genes required for proli

150 Injection of Cas9-guide RNA-lipid complexes targeting the Tmc1(Bth) allele

151 C2c1 is a newly identified guide RNA-mediated type V-B CRISPR-Cas endonuclease that

152 ion in human prostate cancer cells by single guided RNA-mediated targeting activated AKT and increase

153 cophaga bullata, inject these eggs with Cas9/guide RNA mixtures, and transfer injected eggs back into

154 s double-stranded DNA sequences specified by guide RNA molecules and flanked by a protospacer adjacen

155 Cas9's ability to be directed by single 'guide RNA' molecules to target nearly any sequence has b

156 We demonstrate that guide RNA pairs generate deletions that are repaired wit

157 ond CRISPR library was made containing three guide RNAs per construct to target 18 putative transport

158 ncRNAs) that is based on a lentiviral paired-guide RNA (pgRNA) library.

159 than others, and we provide a compendium of guide RNAs predicted to have high efficacy in diverse pa

160 anded target sequences using 5'-hydroxylated guide RNAs rather than the 5'-phosphorylated guides used

161 ructure of a PIWI-clade protein displaying a guide RNA, ready for action.

162 Guide RNA resistant cDNA rescue was a legitimate strateg

163 Crystal structures of Cas9 bound to single-guide RNA reveal a conformation distinct from both the a

164 ng in human CD4(+) T cells using Cas9:single-guide RNA ribonucleoproteins (Cas9 RNPs).

165 sing electroporation of Cas9 nuclease/single-guide RNA ribonucleoproteins and taking advantage of a s

166 ly determined by sequencing the incorporated guide RNA(s) in the primary transgenic events.

167 recruitment of VP64 by dCas9 and a modified guide RNA scaffold gRNA2.0 (designated CRISPR-Act2.0) yi

168 ISPR experiments by presenting ten different guide RNA scoring functions in one simple graphical inte

169 ments revealed that single mismatches in the guide RNA seed sequence reduce the target residence time

170 can be integrated with empirical methods for guide RNA selection into a framework for designing CRISP

171 dly progressing, with marked improvements in guide RNA selection, protein and guide engineering, nove

172 ogrammed to target new sites by altering its guide RNA sequence, and its development as a tool has ma

173 iting technique depends on the choice of the guide RNA sequence, which is facilitated by various webs

174 transactivation domains can act as a potent guide RNA sequence-directed inducer or repressor of gene

175 There exist a number of tools to design the guide RNA sequences and predict potential off-target sit

176 o reveal potential methodologies to engineer guide RNA sequences with improved specificity by conside

177 and gene regulation to predict how changing guide RNA sequences, DNA superhelical densities, Cas9 an

178 genome in a manner that is dependent on the guide RNA sequences.

179 methodology to simultaneously assess single guide RNA (sgRNA) activity across approximately 1,400 ge

180 tructures of SaCas9 in complex with a single guide RNA (sgRNA) and its double-stranded DNA targets, c

181 ge or up to 13 mismatches between the single guide RNA (sgRNA) and its genomic target, which refines

182 occus pyogenes Cas9 alone or bound to single-guide RNA (sgRNA) and target DNA revealed a bilobed prot

183 ome-engineering tool that relies on a single guide RNA (sgRNA) and the Cas9 enzyme for genome editing

184 vity of 12 de novo-designed single synthetic guide RNA (sgRNA) constructs, and found their cleavage e

185 Using hepatic single guide RNA (sgRNA) delivery, we targeted large gene sets

186 In this system, a single guide RNA (sgRNA) directs the endonuclease Cas9 to a tar

187 sequence features that contribute to single guide RNA (sgRNA) efficiency in CRISPR-based screens.

188 ined protein (Cas9) and an engineered single guide RNA (sgRNA) genome editing platform that offers re

189 coccus pyogenes (spCas9) along with a single guide RNA (sgRNA) has emerged as a versatile toolbox for

190 tagged Cas9 and lentivirus encoding a single guide RNA (sgRNA) in primary human lung microvascular EC

191 shold distance from PAM into the Cas9/single-guide RNA (sgRNA) interior is hindered.

192 by microinjection of Cas9 DNA/RNA and single guide RNA (sgRNA) into zygotes to generate modified anim

193 ion of a mixture of Cas9 DNA/mRNA and single-guide RNA (sgRNA) into zygotes.

194 neously delivers the Cas9 protein and single guide RNA (sgRNA) is based on DNA nanoclews, yarn-like D

195 Single-guide RNA (sgRNA) is one of the two key components of th

196 egy for cloning and sequencing paired single guide RNA (sgRNA) libraries and a robust statistical sco

197 platform is simplified by a synthetic single-guide RNA (sgRNA) mimicking the natural dual trans-activ

198 The single guide RNA (sgRNA) of the system recognizes its target se

199 ead (d) Cas9 combined with engineered single guide RNA (sgRNA) scaffolds that bind sets of fluorescen

200 We describe a cloning-free single-guide RNA (sgRNA) synthesis, coupled with streamlined mu

201 luding target selection; cloning-free single-guide RNA (sgRNA) synthesis; microinjection; validation

202 9 activation complexes to investigate single-guide RNA (sgRNA) targeting rules for effective transcri

203 off-target sites in vitro, we used a single guide RNA (sgRNA) that has been previously shown to effi

204 ither multiple cleavages induced by a single-guide RNA (sgRNA) that targets multiple chromosome-speci

205 -Associated Virus 9 (AAV9) to deliver single-guide RNA (sgRNA) that targets the Myh6 locus exclusivel

206 t Palindromic Repeats system allows a single guide RNA (sgRNA) to direct a protein with combined heli

207 e CRISPR-Cas9 system utilizes a short single guide RNA (sgRNA) to direct the endonuclease Cas9 to vir

208 The assembly of single guide RNA (sgRNA) with the Cas9 protein may limit the Ca

209 A at a sequence programmed by a short single-guide RNA (sgRNA), can result in off-target DNA modifica

210 y simply altering the sequence of the single-guide RNA (sgRNA), one can reprogram Cas9 to target diff

211 dCas9) protein assembled with various single-guide RNA (sgRNA), we demonstrated rapid and robust labe

212 (SBH) structure at the 5' end of the single guide RNA (sgRNA), which abrogates the function of CRISP

213 st bioinformatics tools for design of single guide RNA (sgRNA), which determines the efficacy and spe

214 le cut HDR donor, which is flanked by single guide RNA (sgRNA)-PAM sequences and is released after CR

215 ramming the sequence of an associated single guide RNA (sgRNA).

216 expressing an asRNA that sequesters a small guide RNA (sgRNA).

217 a purified protein in complex with a single guide RNA (sgRNA).

218 hat are complementary to a programmed single guide RNA (sgRNA).

219 ormance still relies on well-designed single guide RNAs (sgRNA).

220 (CRISPR)-based genetic screens using single-guide-RNA (sgRNA) libraries have proven powerful to iden

221 f Cas9-encoding mRNA and multiplexing single guide RNAs (sgRNAs) allowed for phenocopy of known mutan

222 n of specific DNA sequences by CRISPR single-guide RNAs (sgRNAs) and fluorescent-protein-fused cataly

223 suffer from interference between the single-guide RNAs (sgRNAs) and from limited gene targeting acti

224 rously predicts off-target binding of single-guide RNAs (sgRNAs) and TALENs.

225 the relative abundance of integrated single guide RNAs (sgRNAs) between populations, which does not

226 Here, we show that some single-guide RNAs (sgRNAs) can induce exon skipping or large ge

227 en, and up to twenty-four multiplexed single guide RNAs (sgRNAs) can induce mutations in 90% of the m

228 e immune cells, we used high-fidelity single guide RNAs (sgRNAs) designed with an sgRNA design tool (

229 Cas9-fluorescent proteins and cognate single-guide RNAs (sgRNAs) efficiently labeled several target l

230 t chemical alterations to synthesized single guide RNAs (sgRNAs) enhance genome editing efficiency in

231 Variants of single-guide RNAs (sgRNAs) for four endogenous loci were used a

232 ess to a database of over 3.4 million single guide RNAs (sgRNAs) for iSTOP (sgSTOPs) targeting 97%-99

233 o accurately predict highly effective single guide RNAs (sgRNAs) for targeting nuclease-dead Cas9-med

234 ed short palindromic repeats (CRISPR) single guide RNAs (sgRNAs) from a single RNA polymerase II or I

235 PR)-based knockout by analysis of 373 single guide RNAs (sgRNAs) in 6 cells lines and show that the o

236 To address this challenge, we design single-guide RNAs (sgRNAs) integrated with up to 16 MS2 binding

237 r, variable activity across different single guide RNAs (sgRNAs) remains a significant limitation.

238 able to wild-type SpCas9 with >85% of single-guide RNAs (sgRNAs) tested in human cells.

239 se applications require the design of single guide RNAs (sgRNAs) that are efficient and specific.

240 in native crRNA:tracrRNA duplexes and single guide RNAs (sgRNAs) that direct Cas9 endonuclease activi

241 r, our understanding of how to select single-guide RNAs (sgRNAs) that mediate efficient Cas9 activity

242 with pools of either single or double single guide RNAs (sgRNAs) to downregulate individual genes or

243 ress transcription in eukaryotic cells using guide RNAs (sgRNAs) to target catalytically inactive Cas

244 ion unless multiple promoter-specific single guide RNAs (sgRNAs) were used.

245 SITE-Seq), using Cas9 programmed with single-guide RNAs (sgRNAs), to identify the sequence of cut sit

246 ripts of viral RNAs (MS2 and PP7) and single-guide RNAs (sgRNAs), which when co-expressed with a clea

247 ng a genome-scale library with 67,405 single-guide RNAs (sgRNAs).

248 igh quantities of biologically active single guide RNAs (sgRNAs).

249 the RNA editing enzyme ADAR to an antisense guide RNA, specific adenosines can be converted to inosi

250 we clone CRISPR/cas9 constructs with single-guide RNAs specifically targeting biogenesis processing

251 eering aimed at altering catalytic function, guide RNA specificity, and PAM requirements and reducing

252 nding and unwinding activity and promiscuous guide RNA specificity.

253 g a minimal recombinase core site flanked by guide RNA-specified sequences.

254 We found that Cas9 is essential for guide RNA stability and that the nuclear Cas9-guide RNA

255 This device consists of a self-targeting guide RNA (stgRNA) that repeatedly directs Streptococcus

256 Guide RNA strands for directed DNA editing by ADAR were

257 ptions of a variety of tools and methods for guide RNA synthesis and mutant identification.

258 yptophan, facilitating zippering up of 20-bp guide RNA:target DNA heteroduplex on ternary complex for

259 Homozygous introduction requires a guide RNA targeting close to the intended mutation, wher

260 cus pyogenes was pre-complexed with a single guide RNA targeting downstream of the ubiquitously expre

261 veloped a CRISPR screen using ~18,000 single guide RNAs targeting >700 kilobases surrounding the gene

262 As a proof-of-concept, we delivered short guide RNAs targeting 3 genes critical for cardiac physio

263 target effects have been reported for CRISPR guide RNAs targeting genes that are amplified at high co

264 silkworms constitutively expressing Cas9 and guide RNAs targeting the BmNPV immediate early-1 (ie-1)

265 ovirus to deliver the nickase Cas9(D10A) and guide RNAs targeting the breakpoint sequences, and anoth

266 the catalytically inactive Cas9 (dCas9) and guide RNAs targeting the endogenous roX locus in the Dro

267 ant Cas9 protein complexed with a library of guide RNAs targeting unwanted species for cleavage, thus

268 we demonstrate that the class 2 type VI RNA-guided RNA-targeting CRISPR-Cas effector Cas13a (previou

269 -Cas immune systems and characterize its RNA-guided, RNA-targeting activity, including regulation by

270 The RNA-guided, RNA-targeting clustered regularly interspaced sh

271 degrade invasive genetic elements by an RNA-guided, RNA-targeting multisubunit interference complex.

272 alindromic repeats (CRISPR)-associated (Cas)-guide RNA technology are reported in maize.

273 n this study demonstrate the utility of Cas9-guide RNA technology as a plant genome editing tool to e

274 h as flexible junctions between helices help guide RNA tertiary folding, the mechanisms through which

275 CRISPR-Cas-mediated genome editing relies on guide RNAs that direct site-specific DNA cleavage facili

276 NAs, we developed a set of aptazyme-embedded guide RNAs that enable small molecule-controlled nucleas

277 enable multiplexed processing and loading of guide RNAs that in turn allow sensitive detection of cel

278 Cells are transfected with two guide RNAs that target respectively HPRT and the gene of

279 s9 (Type II) system binds and cuts DNA using guide RNAs, though the variables that control its on-tar

280 ferase DNMT3A targeted by co-expression of a guide RNA to any 20 bp DNA sequence followed by the NGG

281 able genome engineering (CREATE), links each guide RNA to homologous repair cassettes that both edit

282 n be directed by an engineered RNA-targeting guide RNA to target and inhibit a human +ssRNA virus, he

283 ems use single Cas endonucleases paired with guide RNAs to cleave complementary nucleic acid targets,

284 The CRISPR-Cas9 system uses guide RNAs to direct the Cas9 endonuclease to cleave tar

285 By extending guide RNAs to include effector protein recruitment sites

286 ssed Cas9 together with in vitro transcribed guide-RNAs to introduce engineered 16S rRNA genes.

287 isiae revealed that the Exo9 central channel guides RNA to either Rrp6 or Rrp44 using partially overl

288 s also highly variable, with the majority of guide RNAs unable to generate detectable editing.

289 We identified an efficient OCT4-targeting guide RNA using an inducible human embryonic stem cell-b

290 By first evaluating candidate guide RNAs using a transient embryo assay, we were able

291 Here, we delivered SpCas9 and guide RNAs using adeno-associated viral (AAV) vectors to

292 relative propensities to bind with different guide RNA variants to targeted or off-target sequences.

293 genome, but the efficiency of the predicted guide RNAs varies dramatically.

294 Moreover, we construct a dual guide RNA vector that can make two cuts simultaneously a

295 iously undescribed approach involving single guide RNA, we successfully removed large genome rearrang

296 expression of Cas9 and appropriately chosen guide RNAs, we demonstrate cleavage of cccDNA by Cas9 an

297 elf-cleaving catalytic RNAs (aptazymes) into guide RNAs, we developed a set of aptazyme-embedded guid

298 deficient Cas9 protein and sequence-specific guide RNAs, we show high-resolution and selective isolat

299 coccus pyogenes Cas9 endonuclease and single guide RNAs were cointroduced with or without DNA repair

300 ould be eliminated by reducing the amount of guide RNA with a reduction in on-target editing.