Benjamin Housden

Dr. Benjamin Housden

orcid

Research Focus & Metrics

Academic Career Length

17Years (2010-Present)

(Calculated once Scilit publication data is synced: current year − year of first publication + 1)

Representative Publications

View All Publications
2 March 2015 in Nature Methods
Verified
Journal Article
Open access
PDF file
ACAlejandro Chavez
Alejandro Chavez
JSJonathan Scheiman
Jonathan Scheiman
SVSuhani Vora
Suhani Vora
BPBenjamin W Pruitt
Benjamin W Pruitt
MTMarcelle Tuttle
Marcelle Tuttle
GCGeorge Church
George Church
The fusion of three transcriptional activation domains to a nuclease-deficient Cas9 achieves robust induction of gene expression and can induce differentiation of hiPSCs. The RNA-guided nuclease Cas9 can be reengineered as a programmable transcription factor. However, modest levels of gene activation have limited potential applications. We describe an improved transcriptional regulator obtained through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9. We demonstrate its utility in activating endogenous coding and noncoding genes, targeting several genes simultaneously and stimulating neuronal differentiation of human induced pluripotent stem cells (iPSCs).

23 May 2016 in Nature Methods
Verified
Journal Article
Open access
PDF file
ACAlejandro Chavez
Alejandro Chavez
MTMarcelle Tuttle
Marcelle Tuttle
BPBenjamin W Pruitt
Benjamin W Pruitt
BEBen Ewen-Campen
Ben Ewen-Campen
RCRaj Chari
Raj Chari
GCGeorge Church
George Church
A comparison of seven dCas9-based transcriptional activators shows that VPR, SAM, and Suntag perform best in cell lines from a variety of organisms. Several programmable transcription factors exist based on the versatile Cas9 protein, yet their relative potency and effectiveness across various cell types and species remain unexplored. Here, we compare Cas9 activator systems and examine their ability to induce robust gene expression in several human, mouse, and fly cell lines. We also explore the potential for improved activation through the combination of the most potent activator systems, and we assess the role of cooperativity in maximizing gene expression.

4 November 2013 in Proceedings of the National Academy of Sciences
Verified
Journal Article
Closed access
PDF file
XRXingjie Ren
Xingjie Ren
JSJin Sun
Jin Sun
BHBenjamin Housden
Benjamin Housden
YHYanhui Hu
Yanhui Hu
CRCharles Roesel
Charles Roesel
JNJian-Quan Ni
Jian-Quan Ni
Significance: Using the recently introduced Cas9/sgRNA technique, we have developed a method for specifically targeting Drosophila germ-line cells to generate heritable mutant alleles. We have established transgenic lines that stably express Cas9 in the germ line and compared different promoters and scaffolds of sgRNA in terms of their efficiency of mutagenesis. An overall mutagenesis rate of 74.2% was achieved with this optimized system, as determined by the number of mutant progeny out of all progeny screened. We also evaluated the off-targets associated with the method and established a Web-based resource, as well as a searchable, genome-wide database of predicted sgRNAs appropriate for genome engineering in flies. Our results demonstrate that this optimized Cas9/sgRNA system in Drosophila is efficient, specific, and cost-effective and can be readily applied in a semi-high-throughput manner.

26 August 2016 in eLife
Verified
Journal Article
Open access
HWHuajin Wang
Huajin Wang
MBMichel Becuwe
Michel Becuwe
BHBenjamin Housden
Benjamin Housden
CCChandramohan Chitraju
Chandramohan Chitraju
APAshley J Porras
Ashley J Porras
RFRobert V Farese
Robert V Farese
How proteins control the biogenesis of cellular lipid droplets (LDs) is poorly understood. Using Drosophila and human cells, we show here that seipin, an ER protein implicated in LD biology, mediates a discrete step in LD formation—the conversion of small, nascent LDs to larger, mature LDs. Seipin forms discrete and dynamic foci in the ER that interact with nascent LDs to enable their growth. In the absence of seipin, numerous small, nascent LDs accumulate near the ER and most often fail to grow. Those that do grow prematurely acquire lipid synthesis enzymes and undergo expansion, eventually leading to the giant LDs characteristic of seipin deficiency. Our studies identify a discrete step of LD formation, namely the conversion of nascent LDs to mature LDs, and define a molecular role for seipin in this process, most likely by acting at ER-LD contact sites to enable lipid transfer to nascent LDs.

22 March 2018 in eLife
Verified
Journal Article
Open access
PLPei-Tseng Lee
Pei-Tseng Lee
JZJonathan Zirin
Jonathan Zirin
OKOguz Kanca
Oguz Kanca
WLWen-Wen Lin
Wen-Wen Lin
KSKaren L Schulze
Karen L Schulze
HBHugo J Bellen
Hugo J Bellen
We generated a library of ~1000 Drosophila stocks in which we inserted a construct in the intron of genes allowing expression of GAL4 under control of endogenous promoters while arresting transcription with a polyadenylation signal 3’ of the GAL4. This allows numerous applications. First, ~90% of insertions in essential genes cause a severe loss-of-function phenotype, an effective way to mutagenize genes. Interestingly, 12/14 chromosomes engineered through CRISPR do not carry second-site lethal mutations. Second, 26/36 (70%) of lethal insertions tested are rescued with a single UAS-cDNA construct. Third, loss-of-function phenotypes associated with many GAL4 insertions can be reverted by excision with UAS-flippase. Fourth, GAL4 driven UAS-GFP/RFP reports tissue and cell-type specificity of gene expression with high sensitivity. We report the expression of hundreds of genes not previously reported. Finally, inserted cassettes can be replaced with GFP or any DNA. These stocks comprise a powerful resource for assessing gene function.

31 October 2016 in Nature Reviews Genetics
Verified
Journal Article
Closed access
PDF file
BHBenjamin Housden
Benjamin Housden
MMMatthias Muhar
Matthias Muhar
MGMatthew Gemberling
Matthew Gemberling
CGCharles A. Gersbach
Charles A. Gersbach
DSDidier Y. R. Stainier
Didier Y. R. Stainier
NPNorbert Perrimon
Norbert Perrimon