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David R. Liu
Harvard University
335Publications
70H-index
20.9kCitations
Publications 335
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#1Jeffrey L. Bessen (Broad Institute)H-Index: 2
#2Lena K. Afeyan (Broad Institute)
Last.David R. Liu (Broad Institute)H-Index: 70
view all 8 authors...
The development of site-specific recombinases (SSRs) as genome editing agents is limited by the difficulty of altering their native DNA specificities. Here we describe Rec-seq, a method for revealing the DNA specificity determinants and potential off-target substrates of SSRs in a comprehensive and unbiased manner. We applied Rec-seq to characterize the DNA specificity determinants of several natural and evolved SSRs including Cre, evolved variants of Cre, and other SSR family members. Rec-seq p...
#1Hye-Kyung Lee (NIH: National Institutes of Health)H-Index: 13
#2Michaela Willi (NIH: National Institutes of Health)H-Index: 6
Last.Lothar Hennighausen (NIH: National Institutes of Health)H-Index: 88
view all 7 authors...
A particular challenge in genome engineering has been the simultaneous introduction of mutations into linked (located on the same chromosome) loci. Although CRISPR/Cas9 has been widely used to mutate individual sites, its application in simultaneously targeting of linked loci is limited as multiple nearby double-stranded DNA breaks created by Cas9 routinely result in the deletion of sequences between the cleavage sites. Base editing is a newer form of genome editing that directly converts C∙G-to...
#1Y. Bill Kim (Broad Institute)H-Index: 2
#2Kevin Zhao (Broad Institute)H-Index: 4
Last.David R. Liu (Broad Institute)H-Index: 70
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Delivery into mammalian cells remains a significant challenge for many applications of proteins as research tools and therapeutics. We recently reported that the fusion of cargo proteins to a supernegatively charged (–30)GFP enhances encapsulation by cationic lipids and delivery into mammalian cells. To discover polyanionic proteins with optimal delivery properties, we evaluate negatively charged natural human proteins for their ability to deliver proteins into cultured mammalian cells and human...
#1Holly A. Rees (Broad Institute)H-Index: 8
#2Wei-Hsi YehH-Index: 3
Last.David R. Liu (Broad Institute)H-Index: 70
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In mammalian cells, double-stranded DNA breaks (DSBs) are preferentially repaired through end-joining processes that generally lead to mixtures of insertions and deletions (indels) or other rearrangements at the cleavage site. In the presence of homologous DNA, homology-directed repair (HDR) can generate specific mutations, albeit typically with modest efficiency and a low ratio of HDR products:indels. Here, we develop hRad51 mutants fused to Cas9(D10A) nickase (RDN) that mediate HDR while minim...
#1Andrew V. Anzalone (Broad Institute)
#2Peyton B. Randolph (Broad Institute)
Last.Aditya Raguram (Broad Institute)H-Index: 2
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Most genetic variants that contribute to disease1 are challenging to correct efficiently and without excess byproducts2–5. Here we describe prime editing, a versatile and precise genome editing method that directly writes new genetic information into a specified DNA site using a catalytically impaired Cas9 fused to an engineered reverse transcriptase, programmed with a prime editing guide RNA (pegRNA) that both specifies the target site and encodes the desired edit. We performed more than 175 ed...
#1Sophia C. Kamran (Harvard University)H-Index: 10
#2Jochen K. Lennerz (Harvard University)H-Index: 29
Last.Scott L. Carter (Harvard University)H-Index: 60
view all 14 authors...
Purpose: Molecular properties associated with complete response or acquired resistance to concurrent chemotherapy and radiation therapy (CRT) are incompletely characterized. Experimental Design: We performed integrated whole exome/transcriptome sequencing and immune infiltrate analysis on rectal adenocarcinoma tumors prior to neoadjuvant CRT (pre-CRT) and at time of resection (post-CRT) in 17 patients (8 complete/partial responders [R], 9 nonresponders [NR]). Results: CRT was not associated with...
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