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System Biosciences

2 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Packaging HIV- or FIV-based Lentivector Expression Constructs & Transduction of VSV-G Pseudotyped Viral Particles

Amy Mendenhall¹, Jacob Lesnik¹, Chandreyee Mukherjee¹, Travis Antes¹, Ranjita Sengupta¹

Lentiviral expression vectors are the most effective vehicles for stably expressing different effector molecules or reporter constructs in dividing and non-dividing mammalian cells and whole organisms. Here we provide a protocol on how to package lentivector expression constructs in pseudoviral particles and to transduce target cells using the pseudoviral particles.

Education

CRISPR-Mediated Reorganization of Chromatin Loop Structure

Stefanie L. Morgan^*1,2, Erin Y. Chang^*1, Natasha C. Mariano¹, Abel Bermudez³, Nicole L. Arruda⁴, Fanting Wu⁵, Yunhai Luo¹, Gautam Shankar¹, Star K. Huynh¹, Chiao-Chain Huang⁵, Sharon J. Pitteri³, Kevin C. Wang^1,2,6

¹Department of Dermatology, Program in Epithelial Biology, Stanford University School of Medicine, ²Program in Cancer Biology, Stanford University School of Medicine, ³Canary Center for Cancer Early Detection, Department of Radiology, Stanford University School of Medicine, ⁴Department of Biology, Bridgewater State University, ⁵System Biosciences, ⁶Veterans Affairs Healthcare System

Chromatin looping plays a significant role in gene regulation; however, there have been no technological advances that allow for selective and reversible modification of chromatin loops. Here we describe a powerful system for chromatin loop re-organization using CRISPR-dCas9 (CLOuD9), demonstrated to selectively and reversibly modulate gene expression at targeted loci.