Science 339, 819 (2013); DOI: 10.1126/science.1231143 This copy is for your personal, non-commercial use only. /CropBox [0 0 594 756] << /StructParents 0 /Filter /FlateDecode endobj endobj /Thumb 79 0 R /CreationDate (D:20130922130359-07'00') endobj /Thumb 82 0 R >> The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage. endobj We engineered two different type II CRISPR/Cas systems and demonstrate that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells. The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage. endobj /Parent 3 0 R /MediaBox [0.0 0.0 612.0 792.0] American Association for the Advancement of Science (AAAS). /ExtGState << >> Creative Commons Attribution-Noncommercial-Share Alike. Multiplex genome engineering using CRISPR/Cas systems January 2013 Science 339(6121) DOI: 10.1126/science.1231143 Source PubMed Authors: Le … We engineered two different type II CRISPR systems and demonstrate that Cas9 nucleases can be … /Type /Page We engineered two different type II CRISPR/Cas systems and demonstrate that Cas9 … /T1_0 61 0 R /Type /XObject Cas9 can also be converted into a nicking enzyme to facilitate homology-directed repair with minimal mutagenic activity. 2013-09-22T13:03:59-07:00 << /Type /Metadata 10.1126/science.1229223 Multiplex Genome Engineering Using CRISPR/Cas Systems Le Cong,1,2* F. Ann Ran,1,4* David Cox,1,3 Shuailiang Lin,1,5 Robert Barretto,6 Naomi Habib,1 Patrick D. Hsu,1,4 Xuebing Wu,7 8 8 1† /ModDate (D:20130922130359-07'00') /Type /Page /G11 13 0 R endobj >> /T1_2 63 0 R endstream %PDF-1.4 >> >> << uuid:eaa821b5-1dd1-11b2-0a00-0000488dc9ff /ProcSet [/PDF /Text /ImageC] << /ProcSet [/PDF /Text /ImageB /ImageC /ImageI] /Contents 83 0 R /G3 12 0 R /Contents [86 0 R 87 0 R] /Parent 3 0 R << /Im1 66 0 R /Rotate 0 Massachusetts Institute of Technology. 819-823. /ca .2 /T1_1 62 0 R /X8 15 0 R /Contents 45 0 R %���� /Subtype /Image /X10 16 0 R 2013-09-22T13:03:59-07:00 /Count 7 10 0 obj >> /Pages 3 0 R << >> /F4 20 0 R 12 0 obj /Type /Page >> /Resources 81 0 R endobj endobj Functional elucidation of causal genetic variants and elements requires precise genome editing technologies. << 2013-09-22T13:03:59-07:00 11 0 obj endobj >> In general, the system generated detectable mutations at a frequency of 50–89% for a single locus and 68–74% for double loci in plants ( Supplemental Table 2 ). >> Functional elucidation of causal genetic variants and elements requires precise genome editing technologies. /Rotate 0 �c(6�5)f;��j�mki�ұE}��M?Kx��[k��}f�J�'� ��1hV޳�.6��6���"�X�:���7Q��D��9��\���cDTik��3��-�#�Q��7�o�[�G�!�Ў[G�%�$py��J;��n�}��j�-�#�Q���~��!�U�Џ. /F6 22 0 R /ca 1 /Annots [90 0 R] /Producer (Adobe PDF Library 9.1) /Resources 88 0 R /Contents [67 0 R 68 0 R 69 0 R 70 0 R 71 0 R 72 0 R 73 0 R 74 0 R] /BM /Normal /Thumb 89 0 R << Multiplex Genome Engineering Using CRISPR/Cas Systems Le Cong, 1,2 * F. Ann Ran, 1,4 * David Cox, 1,3 Shuailiang Lin, 1,5 Robert Barretto, 6 Naomi Habib, 1 Patrick D. Hsu, 1,4 Xuebing Wu, 7 Wenyan Jiang, 8 Luciano A. /Metadata 2 0 R 44 0 R] /Annots [24 0 R 25 0 R 26 0 R 27 0 R 28 0 R 29 0 R 30 0 R 31 0 R 32 0 R 33 0 R /Type /Page /MediaBox [0 0 594 756] >> /BM /Normal /F5 21 0 R endobj << /Thumb 76 0 R /ML 4 /Contents 60 0 R Multiplex Genome Engineering Using CRISPR/Cas Systems Le Cong 1 , F Ann Ran, David Cox, Shuailiang Lin, Robert ... Multiplex Genome Engineering Using CRISPR/Cas Systems ... van der Oost J. van der Oost J. >> /Type /Page /CropBox [0 0 594 756] DOI: 10.1126/science.1231143 Science 339, 819 (2013); Le Cong et al. /Rotate 0 /X15 19 0 R 2013 Feb 15;339(6121):819-23. doi: 10.1126/science.1231143. /BitsPerComponent 8 stream 15 0 obj Please share how this access benefits you. endobj �[2{��o �O}�����m�glۣ�M�% 8�X�����^h?\mm ��&*���Dj��o]fGJy}�֥����W.�� x��yp��}�h i�f&i2�2S�4d�IҤM�I�v��1M�6�N2iC�M�dhJ��Ʒ�|��`cc|b�ԧ$�>�u�CƦHZ�m�p��:˫߻Z��~�Ϯ���}w�w?�~��_�R Ѕ7����������f���������R�:f���z����\i�t�*\�n��]߸�Kw�7�Q޺63�p�:r�d�����k�-�U����t���!�z��1�l�k�k��͍h�mlm=�N���.l��6�k��j���ce�p��p�� ? [Le Cong, F Ann Ran, David Cox, Shuailiang Lin, Robert Barretto, Naomi Habib, Patrick D Hsu, Xuebing Wu, Wenyan Jiang /LJ 0 << >> /Length 1241 9 0 obj Lastly, multiple guide sequences can be encoded into a single CRISPR array to enable simultaneous editing of several sites within the mammalian genome, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology. /Resources 75 0 R /X13 17 0 R We engineered two different type II CRISPR/Cas systems and demonstrate that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells. /Type /Page Functional elucidation of causal genetic variants and elements requires precise genome editing technologies. Multiplex genome engineering using CRISPR/Cas systems Science.